Imidazolidinedione derivatives

ABSTRACT

The invention provides a compound of formula (Ia), and pharmaceutically acceptable salts thereof. The invention also provides use of the compounds or salts as modulators of Kv3.1 and/or Kv3.2, and in the treatment of diseases or disorders where a modulator of Kv3.1 and/or Kv3.2 is required, such as depression and mood disorders, hearing disorders, schizophrenia, substance abuse disorders, sleep disorders or epilepsy.

This application is a continuation of U.S. application Ser. No.13/515,097 (U.S. Pat. No. 8,722,695 issued May 13, 2014), filed Jun. 11,2012 (published as US 2012-0289526 A1), which is a U.S. national phaseof International Application No. PCT/EP2010/068946 filed 6 Dec. 2010,which designated the U.S. and claims priority to GB 0921760.5 filed 11Dec. 2009 and GB 1012924.5 filed 30 Jul. 2010, the entire contents ofeach of which are hereby incorporated by reference.

This invention relates to novel compounds, pharmaceutical compositionscontaining them and their use in therapy, in particular as antipsychoticagents.

BACKGROUND TO THE INVENTION

The Kv3 voltage-gated potassium channel family includes four members,Kv3.1, Kv3.2, KV3.3, and Kv3.4. Genes for each of these subtypes cangenerate multiple isoforms by alternative splicing, producing versionswith different C-terminal domains. Thirteen isoforms have beenidentified in mammals to date, but the currents expressed by thesevariants appear identical (Rudy and McBain, 2001, Trends inNeurosciences 24, 517-526). Kv3 channels are activated by depolarizationof the plasma membrane to voltages more positive than −20 mV;furthermore, the channels deactivate rapidly upon repolarization of themembrane. These biophysical properties ensure that the channels opentowards the peak of the depolarizing phase of the neuronal actionpotential to initiate repolarization. Rapid termination of the actionpotential mediated by Kv3 channels allows the neuron to recover morequickly to reach sub-threshold membrane potentials from which furtheraction potentials can be triggered. As a result, the presence of Kv3channels in certain neurons contributes to their ability to fire at highfrequencies (Rudy and McBain, 2001, Trends in Neurosci. 24, 517-526).Kv3.1-3 subtypes are predominant in the CNS, whereas Kv3.4 channels arefound predominantly in skeletal muscle and sympathetic neurons (Weiseret al., 1994, J. Neurosci. 14, 949-972). Kv3.1-3 channel subtypes aredifferentially expressed by sub-classes of interneurons in cortical andhippocampal brain areas (e.g. Chow et al., 1999, J. Neurosci. 19,9332-9345; Martina et al., 1998, J. Neurosci. 18, 8111-8125; McDonaldand Mascagni, 2006, Neurosci. 138, 537-547, Chang et al., 2007, J. Comp.Neurol. 502, 953-972), in the thalamus (e.g. Kasten et al., 2007, J.Physiol. 584, 565-582), cerebellum (e.g. Sacco et al., 2006, Mol. Cell.Neurosci. 33, 170-179), and auditory brain stem nuclei (Li et al., 2001,J. Comp. Neurol. 437, 196-218).

Characterisation of mice in which one or more of the Kv3 subtypes hasbeen deleted shows that the absence of Kv3.1 gives rise to increasedlocomotor activity, altered electroencephalographic activity, and afragmented sleep pattern (Joho et al., 1999, J. Neurophysiol. 82,1855-1864). The deletion of Kv3.2 leads to a reduction in seizurethreshold and altered cortical electroencephalographic activity (Lau etal., 2000, J. Neurosci. 20, 9071-9085). Deletion of Kv3.3 is associatedwith mild ataxia and motor deficits (McMahon et al., 2004, Eur. J.Neurosci. 19, 3317-3327). Double deletion of Kv3.1 and Kv3.3 gives riseto a severe phenotype characterised by spontaneous seizures, ataxia, andan increased sensitivity to the effects of ethanol (Espinosa et al.,2001, J. Neurosci. 21, 6657-6665; Espinosa et al., 2008, J. Neurosci.28, 5570-5581).

The known pharmacology of Kv3 channels is limited. Tetraethylammonium(TEA) has been shown to inhibit the channels at low millimolarconcentrations (Rudy and McBain, 2001, Trends in Neurosci. 24, 517-526),and blood-depressing substance (BDS) toxins from the sea anemone,Anemonia sulcata (Diochot et al., 1998, J. Biol. Chem. 273, 6744-6749),have been shown to selectively inhibit Kv3 channels with high affinity(Yeung et al., 2005, J. Neurosci. 25, 8735-8745). In addition tocompounds acting directly on Kv3 channels, agonists of receptors thatactivate protein kinase A (PKA) and protein kinase C (PKC) have beenshown to modulate Kv3-mediated currents in specific brain areas, leadingto a reduction in the ability of the neurons to fire at high frequency(Atzori et al., 2000, Nat. Neurosci. 3, 791-798; Song et al., 2005, Nat.Neurosci. 8, 1335-1342); these studies suggest that PKA and PKC canspecifically phosphorylate Kv3 channels in a neuron-specific manner,causing a reduction in Kv3-mediated currents. There are no descriptionsin the literature of compounds or biochemical mechanisms that positivelymodulate or activate Kv3 channels.

Bipolar disorder, schizophrenia, anxiety, and epilepsy are seriousdisorders of the central nervous system that have been associated withreduced function of inhibitory interneurons and gamma-amino butyric acid(GABA) transmission (Reynolds et al., 2004, Neurotox. Res. 6, 57-61;Benes et al., 2008, PNAS, 105, 20935-20940; Brambilla et al., 2003, Mol.Psychiatry. 8, 721-37, 715; Aroniadou-Anderjaska et al., 2007, AminoAcids 32, 305-315; Ben-Ari, 2006, Crit. Rev. Neurobiol. 18, 135-144).Parvalbumin positive basket cells that express Kv3 channels in thecortex and hippocampus play a key role in generating feedback inhibitionwithin local circuits (Markram et al., 2004, Nat. Rev. Neurosci. 5,793-807). Given the relative dominance of excitatory synaptic input overinhibitory input to glutamatergic pyramidal neurons in these circuits,fast-firing of interneurons supplying inhibitory input is essential toensure balanced inhibition. Furthermore, accurate timing of inhibitoryinput is necessary to sustain network synchronisation, for example, inthe generation of gamma frequency field potential oscillations that havebeen associated with cognitive function (Fisahn et al., 2005, J. Physiol562, 65-72; Engel et al., 2001, Nat. Rev. Neurosci. 2, 704-716).Notably, a reduction in gamma oscillations has been observed in patientswith schizophrenia (Spencer et al., 2004, PNAS 101, 17288-17293).Consequently, positive modulators of Kv3 channels might be expected toenhance the firing capabilities of specific groups of fast-firingneurons in the brain. These effects may be beneficial in disordersassociated with abnormal activity of these neuronal groups.

In addition, Kv3.2 channels have been shown to be expressed by neuronsof the superchiasmatic nucleus (SCN) the main circadian pacemaker in theCNS (Schulz and Steimer, 2009, CNS Drugs 23 Suppl 2, 3-13). We haveshown that the expression of Kv3.2 channels varies over a 24 hourperiod; thus Kv3.2 channel expression may contribute to changes in thefiring properties of neurons in the SCN and thus influence circadianrhythm. Consequently, drugs that modulate the activity of Kv3.2 channelscould influence circadian rhythm and thus be useful in the treatment ofrelated disorders.

Hearing loss represents an epidemic that affects approximately 16% ofthe population in Europe and the US (Goldman and Holme, 2010, DrugDiscovery Today 15, 253-255), with a prevalence estimated at 250 millionpeople worldwide (B. Shield, 2006, Evaluation of the social and economiccosts of hearing impairment. A report for Hear-It AISBL:www.hear-it.org/multimedia/Hear It Report October 2006.pdf). As lifeexpectancy continues to increase, so too will the number of peoplesuffering from hearing disorders. Furthermore, it is believed thatmodern lifestyles may exacerbate this burden as the younger generationages. Hearing conditions, including tinnitus have a profound effect onthe quality of life, causing social isolation, depression, work andrelationship difficulties, low self-esteem, and prejudice. Voltage-gatedion channels of the Kv3 family are expressed at high levels in auditorybrainstem nuclei (Li et al., 2001, J. Comp. Neurol. 437, 196-218) wherethey permit the fast firing of neurons that transmit auditoryinformation from the cochlear to higher brain regions. Loss of Kv3.1channel expression in central auditory neurons is observed in hearingimpaired mice (von Hehn et al., 2004, J. Neurosci. 24, 1936-1940), and adecline in Kv3.1 expression may be associated with loss of hearing inaged mice (Jung et al. 2005 Neurol. Res. 27, 436-440). Furthermore,pathological plasticity of auditory brainstem networks is likely tocontribute to symptoms of tinnitus that are experienced by many peoplesuffering from hearing loss of different types. Recent studies haveshown that regulation of Kv3.1 channel function and expression has amajor role in controlling auditory neuron excitability (Kaczmarek etal., 2005, Hearing Res. 206, 133-145), suggesting that this mechanismcould account for some of the plastic changes that give rise totinnitus. Finally, Fraglie X syndrome and autism are frequentlyassociated with hypersensitivity to sensory input, including auditorystimuli. Recent findings suggest that the protein coded by the FMR-Igene, whose mutation or absence gives rise to Fragile X syndrome, maydirectly regulate the expression of Kv3.1 channels in the auditorybrainstem nuclei (Strumbos et al., 2010, J. Neuroscience, in press),suggesting that mis-regulation of Kv3.1 channels could give rise tohyperacusis in patients suffering from Fragile X or autism.Consequently, we propose that small molecule modulators of Kv3 channelsin auditory brainstem nuclei could have a benefit in the treatment ofdisorders of hearing, including tinnitus and auditory hyper-acuityassociated with Fragile X syndrome and autism.

In a first aspect therefore, the invention provides a compound offormula (Ia)

wherein:

R¹ is halo, C₁₋₄ alkyl, C₁₋₄ alkoxy, halo-C₁₋₄alkyl, halo-C₁₋₄alkoxy, orcyano;

R² is H, halo, cyano, C₁₋₄ alkyl or C₁₋₄ alkoxy; with the proviso thatwhen R₂ is H, R₁ is not in the para position;

X is C or N;

Y is C or N;

R³ is C₁₋₄ alkyl;

R⁴ is H, deuterium, or C₁₋₄alkyl; or R₃ and R₄ can be fused to form aC₃₋₄ spiro carbocyclyl group;

or a pharmaceutically acceptable salt thereof.

In a second aspect the invention provides a compound of formula (Ib)

wherein:

R¹ is halo, C₁₋₄ alkyl or C₁₋₄ alkoxy, halo-C₁₋₄alkoxy, cyano;

R² is H, halo, C₁₋₄ alkyl and C₁₋₄ alkoxy; with the proviso that when R₂is H, R₁ is not in the para position;

X is C or N;

Y is C or N;

R³ is C₁₋₄ alkyl;

R⁴ is H, deuterium, C₁₋₄alkyl; or R₃ and R₄ can be fused to form a C₃₋₄spiro carbocycly group;

or a pharmaceutically acceptable salt thereof.

In a third aspect therefore, the invention provides a compound offormula (Ic)

wherein:

R¹ is halo, C₁₋₄ alkyl or C₁₋₄ alkoxy;

R² is H, halo, C₁₋₄ alkyl and C₁₋₄ alkoxy; with the proviso that when R₂is H, R₁ is not in the para position;

X is C or N;

Y is C or N;

R³ is C₁₋₄ alkyl;

R⁴ is H, deuterium, C₁₋₄alkyl; or R₃ and R₄ can be fused to form a C₃₋₄spiro carbocycly group;

or a pharmaceutically acceptable salt thereof.

As used herein below, “Formula (I)” means any one of Formula (Ia), (Ib),or (Ic).

In one embodiment of the invention R¹ is C₁₋₄ alkoxy. In anotherembodiment of the invention R¹ is methoxy.

In one embodiment of the invention R¹ is C₁₋₄ alkyl. In anotherembodiment of the invention R¹ is methyl. In a further embodiment of theinvention R¹ is ethyl. In a yet further embodiment of the invention R¹is propyl. In a yet further embodiment of the invention R¹ is butyl.

In one embodiment of the invention R¹ is halo. In another embodiment ofthe invention R¹ is chloro. In a further embodiment of the invention R¹is fluoro.

In one embodiment of the invention R¹ is halo-C₁₋₄alkoxy. In anotherembodiment of the invention R¹ is trifluoromethoxy.

In one embodiment of the invention R¹ is halo-C₁₋₄alkyl. In anotherembodiment of the invention R¹ is trifluoromethyl.

In one embodiment of the invention R¹ is cyano.

In one embodiment of the invention, R² is H.

In one embodiment of the invention R² is C₁₋₄alkyl. In anotherembodiment of the invention R² is methyl.

In one embodiment of the invention R² is halo. In another embodiment ofthe invention, R² is chloro. In a further embodiment of the invention R²is fluoro.

In one embodiment of the invention R² is C₁₋₄ alkyl.

In one embodiment of the invention R² is cyano.

In one embodiment of the invention X is C and Y is C.

In one embodiment of the invention X is N and Y is C.

In one embodiment of the invention X is N and Y is N.

In one embodiment of the invention R³ is methyl. In another embodimentof the invention R³ is ethyl. In a further embodiment of the inventionR³ is propyl. In a yet further embodiment of the invention R³ is butlyl.

In one embodiment of the invention R⁴ is H.

In one embodiment of the invention R⁴ is deuterium.

In one embodiment of the invention R⁴ is C₁₋₄ alkyl. In anotherembodiment of the invention R⁴ is methyl.

In one embodiment of the invention R³ and R⁴ together form a C₃₋₄ spirocarbocyl. In anther embodiment of the invention R³ and R⁴ together forma C₃ spiro carbocycyl. In a further embodiment of the invention R³ andR⁴ together form a C₄ spiro carbocycyl.

In one embodiment of the invention R³ is C₁₋₄ alkyl, R⁴ is H and theabsolute configuration of the stereogenic centre is R.

In one embodiment of the invention R¹ is C₁₋₄alkyl, C₁₋₄ alkoxy, orhalo-C₁₋₄ alkoxy; R² is H, cyano or alkyl; X is N, Y is N or C, R₃ isC₁₋₄ alkyl, and R⁴ is C₁₋₄ alkyl or H; or a pharmaceutically acceptablesalt thereof.

In one embodiment of the invention R₁ is propyl, butyl, methoxy,propoxy, or trifluoromethoxy; R² is H, cyano or methyl; X is N, Y is Nor C, R³ is ethyl, and R⁴ is methyl or H; or a pharmaceuticallyacceptable salt thereof.

In one embodiment of the invention R¹ is methoxy and R² is methyl. Inanother embodiment of the invention R¹ is methoxy in the meta positionand R² is methyl in the para position. In a further embodiment of theinvention R¹ is methoxy in the meta position, R² is methyl in the paraposition, R³ is C₁₋₄ alkyl, R⁴ is H, R³ is in the R configuration. In ayet further embodiment of the invention R¹ is methoxy in the metaposition, R² is methyl in the para position, X is N, Y is C, R³ is C₁₋₄alkyl, R⁴ is H and the absolute configuration of the stereogenic centreis R. In a still further embodiment of the invention R¹ is methoxy inthe meta position, R² is methyl in the para position, X is N, Y is C, R³is ethyl, R⁴ is H and the absolute configuration of the stereogeniccentre is R.

In one embodiment of the invention the compound is selected from thegroup consisting of:

-   (5R)-5-methyl-3-{4-[(3-methylphenyl)oxy]phenyl}-2,4-imidazolidinedione;-   (5R)-5-methyl-3-(4-{[3-(methyloxy)phenyl]oxy}phenyl)-2,4-imidazolidinedione;-   (5R)-3-(4-{[3-(ethyloxy)phenyl]oxy}phenyl)-5-methyl-2,4-imidazolidinedione;-   (5R)-3-{4-[(3-chloro-5-fluorophenyl)oxy]phenyl}-5-methyl-2,4-imidazolidinedione;-   (5R)-3-{4-[(3-chloro-4-fluorophenyl)oxy]phenyl}-5-methyl-2,4-imidazolidinedione;-   (5S)-3-{4-[(3-chloro-4-fluorophenyl)oxy]phenyl}-5-methyl-2,4-imidazolidinedione;-   (5R)-5-methyl-3-(4-{[2-methyl-5-(methyloxy)phenyl]oxy}phenyl)-2,4-imidazolidinedione;-   (5R)-5-methyl-3-(4-{[4-methyl-3-(methyloxy)phenyl]oxy}phenyl)-2,4-imidazolidinedione;-   (5R)-5-methyl-3-(6-{[3-(1-methylethyl)phenyl]oxy}-3-pyridinyl)-2,4-imidazolidinedione;-   (5R)-5-methyl-3-[6-({3-[(1-methylethyl)oxy]phenyl}oxy)-3-pyridinyl]-2,4-imidazolidinedione;-   (5R)-3-{6-[(2,5-dimethylphenyl)oxy]-3-pyridinyl}-5-methyl-2,4-imidazolidinedione;-   (5R)-3-{6-[(2,3-dimethylphenyl)oxy]-3-pyridinyl}-5-methyl-2,4-imidazolidinedione;-   (5R)-3-{6-[(2,6-dimethylphenyl)oxy]-3-pyridinyl}-5-methyl-2,4-imidazolidinedione;-   (5R)-3-{6-[(2-ethylphenyl)oxy]-3-pyridinyl}-5-methyl-2,4-imidazolidinedione;-   (5R)-5-methyl-3-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)-2,4-imidazolidinedione;-   (5R)-5-methyl-3-(6-{[2-methyl-5-(methyloxy)phenyl]oxy}-3-pyridinyl)-2,4-imidazolidinedione;-   (5R)-5-methyl-3-(6-{[2-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)-2,4-imidazolidinedione;-   (5R)-5-ethyl-3-(4-{[3-(methyloxy)phenyl]oxy}phenyl)-2,4-imidazolidinedione;-   (5R)-5-ethyl-3-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)-2,4-imidazolidinedione;-   (5S)-5-ethyl-3-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)-2,4-imidazolidinedione;-   (5R)-5-ethyl-3-(6-{[3-(1-methylethyl)phenyl]oxy}-3-pyridinyl)-2,4-imidazolidinedione;-   5,5-dimethyl-3-(4-{[3-(methyloxy)phenyl]oxy}phenyl)-2,4-imidazolidinedione;-   3-{4-[(2,3-dimethylphenyl)oxy]phenyl}-5,5-dimethyl-2,4-imidazolidinedione;-   3-{6-[(2-ethylphenyl)oxy]-3-pyridinyl}-5,5-dimethyl-2,4-imidazolidinedione;-   3-{6-[(2,6-dimethylphenyl)oxy]-3-pyridinyl}-5,5-dimethyl-2,4-imidazolidinedione;-   (5R)-5-(1-methylethyl)-3-(4-{[4-methyl-3-(methyloxy)phenyl]oxy}phenyl)-2,4-imidazolidinedione;-   (5R)-5-methyl-3-(2-{[3-(1-methylethyl)phenyl]oxy}-5-pyrimidinyl)-2,4-imidazolidinedione;-   (5R)-5-ethyl-3-(2-{[3-(ethyloxy)-4-methylphenyl]oxy}-5-pyrimidinyl)-2,4-imidazolidinedione;-   (5R)-5-(1,1-dimethylethyl)-3-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)-2,4-imidazolidinedione;-   (5R)-5-ethyl-5-methyl-3-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)-2,4-imidazolidinedione;-   7-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)-5,7-diazaspiro[3.4]octane-6,8-dione;-   6-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)-4,6-diazaspiro[2.4]heptane-5,7-dione;-   4-{[5-(4,4-dimethyl-2,5-dioxo-1-imidazolidinyl)-2-pyridinyl]oxy}-2-(1-methylethyl)benzonitrile;-   4-{[5-(4,4-dimethyl-2,5-dioxo-1-imidazolidinyl)-2-pyridinyl]oxy}-2-[(trifluoromethyl)oxy]benzonitrile;-   3-{6-[(4-fluoro-3-methylphenyl)oxy]-3-pyridinyl}-5,5-dimethyl-2,4-imidazolidinedione;-   3-{6-[(4-fluoro-2-methylphenyl)oxy]-3-pyridinyl}-5,5-dimethyl-2,4-imidazolidinedione;-   5,5-dimethyl-3-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)-2,4-imidazolidinedione;-   (5R)-5-(1-methylethyl)-3-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)-2,4-imidazolidinedione;-   3-(6-{[2-(1,1-dimethylethyl)phenyl]oxy}-3-pyridinyl)-5,5-dimethyl-2,4-imidazolidinedione;-   3-(2-{[2-(1,1-dimethylethyl)phenyl]oxy}-5-pyrimidinyl)-5,5-dimethyl-2,4-imidazolidinedione;-   (5R)-5-ethyl-5-methyl-3-(2-{[4-methyl-3-(methyloxy)phenyl]oxy}-5-pyrimidinyl)-2,4-imidazolidinedione;-   (5R)-5-ethyl-3-(2-{[3-(ethyloxy)-4-methylphenyl]oxy}-5-pyrimidinyl)-5-methyl-2,4-imidazolidinedione;-   5,5-dimethyl-3-[6-({3-[(trifluoromethyl)oxy]phenyl}oxy)-3-pyridinyl]-2,4-imidazolidinedione;-   4-{[5-(4,4-dimethyl-2,5-dioxo-1-imidazolidinyl)-2-pyridinyl]oxy}-3-ethylbenzonitrile;-   2-chloro-4-{[5-(4,4-dimethyl-2,5-dioxo-1-imidazolidinyl)-2-pyridinyl]oxy}benzonitrile;-   5,5-dimethyl-3-[6-({4-methyl-3-[(trifluoromethyl)oxy]phenyl}oxy)-3-pyridinyl]-2,4-imidazolidinedione;-   4-{[5-(4,4-dimethyl-2,5-dioxo-1-imidazolidinyl)-2-pyridinyl]oxy}-2-(methyloxy)benzonitrile;-   4-{[5-(4,4-dimethyl-2,5-dioxo-1-imidazolidinyl)-2-pyridinyl]oxy}-3-methylbenzonitrile;-   4-{[5-(4,4-dimethyl-2,5-dioxo-1-imidazolidinyl)-2-pyridinyl]oxy}-3-(trifluoromethyl)benzonitrile;-   4-{[5-(4,4-dimethyl-2,5-dioxo-1-imidazolidinyl)-2-pyridinyl]oxy}-2-ethylbenzonitrile;-   4-{[5-(4,4-dimethyl-2,5-dioxo-1-imidazolidinyl)-2-pyrimidinyl]oxy}-2-ethylbenzonitrile;-   3-cyclopropyl-4-{[5-(4,4-dimethyl-2,5-dioxo-1-imidazolidinyl)-2-pyridinyl]oxy}benzonitrile;-   4-{[5-(4,4-dimethyl-2,5-dioxo-1-imidazolidinyl)-2-pyridinyl]oxy}-3-(1,1-dimethylethyl)benzonitrile;-   2-[(cyclopropylmethyl)oxy]-4-{[5-(4,4-dimethyl-2,5-dioxo-1-imidazolidinyl)-2-pyridinyl]oxy}benzonitrile;-   4-{[5-(4,4-dimethyl-2,5-dioxo-1-imidazolidinyl)-2-pyridinyl]oxy}-2-(ethyloxy)benzonitrile;-   2-cyclopropyl-4-{[5-(4,4-dimethyl-2,5-dioxo-1-imidazolidinyl)-2-pyridinyl]oxy}benzonitrile;-   5,5-dimethyl-3-[2-({4-methyl-3-[(trifluoromethyl)oxy]phenyl}oxy)-5-pyrimidinyl]-2,4-imidazolidinedione;-   4-{[5-(4,4-dimethyl-2,5-dioxo-1-imidazolidinyl)-2-pyrimidinyl]oxy}-3-(1,1-dimethylethyl)benzonitrile;-   4-{[5-(4,4-dimethyl-2,5-dioxo-1-imidazolidinyl)-2-pyridinyl]oxy}-2-[(1-methylethyl)oxy]benzonitrile;-   4-({5-[(4R)-4-ethyl-4-methyl-2,5-dioxo-1-imidazolidinyl]-2-pyridinyl}oxy)-2-[(1-methylethyl)oxy]benzonitrile;-   3-cyclopropyl-4-({5-[(4R)-4-ethyl-4-methyl-2,5-dioxo-1-imidazolidinyl]-2-pyridinyl}oxy)benzonitrile;-   4-({5-[(4R)-4-ethyl-4-methyl-2,5-dioxo-1-imidazolidinyl]-2-pyridinyl}oxy)-2-[(trifluoromethyl)oxy]benzonitrile;-   2-cyclopropyl-4-({5-[(4R)-4-ethyl-4-methyl-2,5-dioxo-1-imidazolidinyl]-2-pyridinyl}oxy)benzonitrile;-   (5R)-5-ethyl-5-methyl-3-[2-({4-methyl-3-[(trifluoromethyl)oxy]phenyl}oxy)-5-pyrimidinyl]-2,4-imidazolidinedione;-   3-(1,1-dimethylethyl)-4-({5-[(4R)-4-ethyl-4-methyl-2,5-dioxo-1-imidazolidinyl]-2-pyrimidinyl}oxy)benzonitrile;-   3-(1,1-dimethylethyl)-4-({5-[(4R)-4-ethyl-4-methyl-2,5-dioxo-1-imidazolidinyl]-2-pyridinyl}oxy)benzonitrile;-   4-{[4-(4,4-dimethyl-2,5-dioxo-1-imidazolidinyl)phenyl]oxy}-2-(methyloxy)benzonitrile;-   4-{[4-(4,4-dimethyl-2,5-dioxo-1-imidazolidinyl)phenyl]oxy}-2-(ethyloxy)benzonitrile;-   4-({4-[(4R)-4-ethyl-2,5-dioxo-1-imidazolidinyl]phenyl}oxy)-2-(ethyloxy)benzonitrile;-   3-cyclopropyl-4-({5-[(4R)-4-ethyl-2,5-dioxo-1-imidazolidinyl]-2-pyridinyl}oxy)benzonitrile;-   3-(1,1-dimethylethyl)-4-({5-[(4R)-4-ethyl-2,5-dioxo-1-imidazolidinyl]-2-pyridinyl}oxy)benzonitrile;-   4-({5-[(4R)-4-ethyl-2,5-dioxo-1-imidazolidinyl]-2-pyridinyl}oxy)-2-(methyloxy)benzonitrile;-   4-({4-[(4R)-4-ethyl-2,5-dioxo-1-imidazolidinyl]phenyl}oxy)-2-(methyloxy)benzonitrile;-   2-[(cyclopropylmethyl)oxy]-4-({5-[(4R)-4-ethyl-2,5-dioxo-1-imidazolidinyl]-2-pyridinyl}oxy)benzonitrile;-   (5R)-5-ethyl-3-[6-({4-methyl-3-[(trifluoromethyl)oxy]phenyl}oxy)-3-pyridinyl]-2,4-imidazolidinedione;-   2-cyclopropyl-4-({5-[(4R)-4-ethyl-2,5-dioxo-1-imidazolidinyl]-2-pyridinyl}oxy)benzonitrile;-   4-({5-[(4R)-4-ethyl-2,5-dioxo-1-imidazolidinyl]-2-pyridinyl}oxy)-2-(1-methylethyl)benzonitrile;-   4-({5-[(4R)-4-ethyl-2,5-dioxo-1-imidazolidinyl]-2-pyridinyl}oxy)-2-(1-methylethyl)benzonitrile;-   (5R)-5-ethyl-3-[2-({4-methyl-3-[(trifluoromethyl)oxy]phenyl}oxy)-5-pyrimidinyl]-2,4-imidazolidinedione;-   4-({5-[(4R)-4-ethyl-2,5-dioxo-1-imidazolidinyl]-2-pyridinyl}oxy)-2-[(1-methylethyl)oxy]benzonitrile;-   4-({5-[(4R)-4-ethyl-2,5-dioxo-1-imidazolidinyl]-2-pyridinyl}oxy)-3-methylbenzonitrile;-   4-({5-[(4R)-4-ethyl-2,5-dioxo-1-imidazolidinyl]-2-pyridinyl}oxy)-2-[(trifluoromethyl)oxy]benzonitrile;-   3-ethyl-4-({5-[(4R)-4-ethyl-2,5-dioxo-1-imidazolidinyl]-2-pyrimidinyl}oxy)benzonitrile;-   4-({5-[(4R)-4-ethyl-2,5-dioxo-1-imidazolidinyl]-2-pyrimidinyl}oxy)-3-methylbenzonitrile;-   3-(1,1-dimethylethyl)-4-({5-[(4R)-4-ethyl-2,5-dioxo-1-imidazolidinyl]-2-pyrimidinyl}oxy)benzonitrile    and-   4-({5-[(4R)-4-ethyl-4-methyl-2,5-dioxo-1-imidazolidinyl]-2-pyridinyl}oxy)-2-(1-methylethyl)benzonitrile    or

a pharmaceutically acceptable salt thereof.

In one embodiment of the invention, the compound is selected from thegroup consisting of:

-   (5R)-5-ethyl-3-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl;-   (5R)-5-ethyl-5-methyl-3-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)-2,4-imidazolidinedione;-   4-({5-[(4R)-4-ethyl-4-methyl-2,5-dioxo-1-imidazolidinyl]-2-pyridinyl}oxy)-2-[(trifluoromethyl)oxy]benzonitrile;-   (5R)-5-ethyl-5-methyl-3-[2-({4-methyl-3-[(trifluoromethyl)oxy]phenyl}oxy)-5-pyrimidinyl]-2,4-imidazolidinedione;-   3-(1,1-dimethylethyl)-4-({5-[(4R)-4-ethyl-4-methyl-2,5-dioxo-1-imidazolidinyl]-2-pyridinyl}oxy)benzonitrile;-   (5R)-5-ethyl-3-[6-({4-methyl-3-[(trifluoromethyl)oxy]phenyl}oxy)-3-pyridinyl]-2,4-imidazolidinedione;-   4-({5-[(4R)-4-ethyl-2,5-dioxo-1-imidazolidinyl]-2-pyridinyl}oxy)-2-(1-methylethyl)benzonitrile;-   4-({5-[(4R)-4-ethyl-2,5-dioxo-1-imidazolidinyl]-2-pyridinyl}oxy)-2-(1-methylethyl)benzonitrile;-   (5R)-5-ethyl-3-[2-({4-methyl-3-[(trifluoromethyl)oxy]phenyl}oxy)-5-pyrimidinyl]-2,4-imidazolidinedione;-   4-({5-[(4R)-4-ethyl-2,5-dioxo-1-imidazolidinyl]-2-pyridinyl}oxy)-2-[(1-methylethyl)oxy]benzonitrile;-   4-({5-[(4R)-4-ethyl-2,5-dioxo-1-imidazolidinyl]-2-pyridinyl}oxy)-2-[(trifluoromethyl)oxy]benzonitrile;-   3-(1,1-dimethylethyl)-4-({5-[(4R)-4-ethyl-2,5-dioxo-1-imidazolidinyl]-2-pyrimidinyl}oxy)benzonitrile;

or a pharmaceutically acceptable salt thereof.

For the avoidance of doubt, the embodiments of any one feature of thecompounds of the invention may be combined with any embodiment ofanother feature of compounds of the invention to create a furtherembodiment.

The term ‘halo’ or ‘halogen’ as used herein, refers to a fluorine,chlorine, bromine or iodine atom.

When the compound contains a (C₁₋₄)alkyl group, whether alone or formingpart of a larger group, e.g. (C₁₋₄)alkoxy, the alkyl group may bestraight chain, branched, cyclic, or a combination thereof. Examples of(C₁₋₄)alkyl are methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl,sec-butyl, tert-butyl, cyclopropyl and cyclobutyl. An example of(C₁₋₄)alkoxy is methoxy. An example of halo-C₁₋₄alky is trifluoromethyl.An example of halo-C₁₋₄alkoxy is trifluoromethoxy.

It will be appreciated that for use in medicine the salts of thecompounds of formula (I) should be pharmaceutically acceptable. Suitablepharmaceutically acceptable salts will be apparent to those skilled inthe art. Pharmaceutically acceptable salts include those described byBerge, Bighley and Monkhouse J. Pharm. Sci (1977) 66, pp 1-19. Suchpharmaceutically acceptable salts include acid addition salts formedwith inorganic acids e.g. hydrochloric, hydrobromic, sulphuric, nitricor phosphoric acid and organic acids e.g. succinic, maleic, acetic,fumaric, citric, tartaric, benzoic, p-toluenesulfonic, methanesulfonicor naphthalenesulfonic acid. Other salts e.g. oxalates or formates, maybe used, for example in the isolation of compounds of formula (I) andare included within the scope of this invention.

Certain of the compounds of formula (I) may form acid addition saltswith one or more equivalents of the acid. The present invention includeswithin its scope all possible stoichiometric and non-stoichiometricforms.

The compounds of formula (I) may be prepared in crystalline ornon-crystalline form and, if crystalline, may optionally be solvated,eg. as the hydrate. This invention includes within its scopestoichiometric solvates (eg. hydrates) as well as compounds containingvariable amounts of solvent (eg. water).

It will be understood that the invention includes pharmaceuticallyacceptable derivatives of compounds of formula (I) and that these areincluded within the scope of the invention.

As used herein “pharmaceutically acceptable derivative” includes anypharmaceutically acceptable ester or salt of such ester of a compound offormula (I) which, upon administration to the recipient is capable ofproviding (directly or indirectly) a compound of formula (I) or anactive metabolite or residue thereof.

It is to be understood that the present invention encompasses allisomers of formula (I) and their pharmaceutically acceptablederivatives, including all geometric, tautomeric and optical forms, andmixtures thereof (e.g. racemic mixtures). Where additional chiralcentres are present in compounds of formula (I), the present inventionincludes within its scope all possible diastereoismers, includingmixtures thereof. The different isomeric forms may be separated orresolved one from the other by conventional methods, or any given isomermay be obtained by conventional synthetic methods or by stereospecificor asymmetric syntheses.

The subject invention also includes isotopically-labeled compounds whichare identical to those recited in formula (I) but for the fact that oneor more atoms are replaced by an atom having an atomic mass or massnumber different from the atomic mass or mass number most commonly foundin nature. Examples of isotopes that can be incorporated into compoundsof the invention include isotopes of hydrogen, carbon, nitrogen, oxygen,fluorine, iodine and chlorine such as ³H, ¹¹C, ¹⁴C, ¹⁸F, ¹²³I or ¹²⁵I.

Compounds of the present invention and pharmaceutically acceptable saltsof said compounds that contain the aforementioned isotopes and/or otherisotopes of other atoms are within the scope of the present invention.Isotopically labeled compounds of the present invention, for examplethose into which radioactive isotopes such as ³H or ¹⁴C have beenincorporated, are useful in drug and/or substrate tissue distributionassays. Tritiated, ie. ³H, and carbon-14, ie. ¹⁴C, isotopes areparticularly preferred for their ease of preparation and detectability.¹¹C and ¹⁸F isotopes are particularly useful in PET (positron emissiontomography).

Since the compounds of formula (I) are intended for use inpharmaceutical compositions it will readily be understood that they areeach preferably provided in substantially pure form, for example atleast 60% pure, more suitably at least 75% pure and preferably at least85%, especially at least 98% pure (% are on a weight for weight basis).Impure preparations of the compounds may be used for preparing the morepure forms used in the pharmaceutical compositions.

According to a further aspect of the present invention there is provideda process for the preparation of compounds of formula (I) andderivatives thereof. The following schemes detail some synthetic routesto compounds of the invention. In the following schemes reactive groupscan be protected with protecting groups and deprotected according towell established techniques.

In general, the compounds of formula (I) may be made according to theorganic synthesis techniques known to those skilled in this field, aswell as by the representative methods set forth in the Examples.

Compounds of formula (I), and salts and solvates thereof, may beprepared by the general methods outlined hereinafter. In the followingdescription, the groups R₁, R₂, X, Y, R₃, R₄ have the meanings aspreviously defined for compounds of formula (I) unless otherwise stated.

step (ii): Compounds of formula (I) can be prepared by cyclization ofcompounds of formula (II) in a solvent e.g. dichloromethane with acarbonylating agent e.g. triphosgene preferentially prediluted in thesame solvent and added in a second time at 0° C. in presence of a basee.g. triethylamine. In some cases ethyl acetate could be used as asolvent. Optionally a catalytic amount of DMAP can be added.

step (i): Compounds of formula (II) can be prepared from compounds offormula (III) by removal of the BOC protective group in acidicconditions e.g. TFA in a solvent e.g. dichloromethane e.g. at 0° C., RT.

step (ii): Compounds of formula (III) can be prepared from anilines offormula (IV) and N-protected amino acids of formula (V) by amidiccoupling in presence of a base e.g. DIPEA and of a coupling agent e.g.HATU, TBTU, HBTU in a solvent such as N,N-dimethylformamide.

step (i): Some N-Boc protected amino acids of formula (V) arecommercially available e.g.N-{[(1,1-dimethylethyl)oxy]carbonyl}-2-methylalanine from for exampleAldrich, N-{[(1,1-dimethylethyl)oxy]carbonyl}-D-alanine from for exampleAldrich, (2R)-2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)butanoic acidfrom for example Bachem UK Ltd,N-{[(1,1-dimethylethyl)oxy]carbonyl}-D-isovaline from for example Nagase& Co Ltd.

N-protected amino acids of formula (V) can also be prepared fromcompounds of formula (VI) for example with Boc-anhydride in presence ofa base e.g. aqueous NaHCO3, aqueous sodium hydroxide in a solvent suchas THF, methanol, dioxane. Many descriptions are available in theliterature (for example Tetrahedron, 2006, 62(42), 9966-9972)

Some anilines (IV) are commercially available, e.g.4-{[3-(methyloxy)phenyl]oxy}aniline for example from ChemBridgeCorporation.

Other anilines can be prepared from the nitro compounds (VII). Suitablereactions conditions to transform (VII) into (IV) are for example:

-   -   reduction in presence of Fe powder and ammonium chloride in a        solvent such as ethanol or a mixture THF/water with heating or        not    -   reduction in presence of Zn powder and ammonium chloride in a        solvent such as ethanol or a mixture THF/water with heating or        not    -   reduction with tin chloride hydrate in a solvent such as ethyl        acetate, ethanol with heating for example at reflux

Anilines of formula (IVa), wherein R¹ is (C₁₋₄ alkyl or C₁₋₄ alkoxy), R²is (H, C₁₋₄ alkyl or C₁₋₄ alkoxy) and (X, Y)≠(N,N) can be prepared fromthe nitro compounds (VIIa) with the conditions described on scheme 3 orwith the following conditions:

-   -   hydrogenation with H₂ with a catalyst such as Pd/C in a solvent        such as methanol, ethanol, THF, a mixture methanol/ethyl acetate        with heating or not    -   reduction with hydrazine hydrate and a catalytic amount of Pd/C        in a solvent such as ethanol with heating

Compounds of formula (VIIb) wherein X═Y═C or (X═C, Y═N) or (X═N, Y═C)can be prepared by nucleophilic aromatic substitution. In this reactionare used a nitro derivative of formula (VIIIb) wherein Z═F (usually when[X═C, Y═C]) or Z═Cl (usually when [X═N, Y═C] or [X═C, Y═N]) and a phenolof formula (IX) in presence of a base in a solvent such as

-   -   potassium carbonate e.g. in N,N-dimethylformamide or in        acetonitrile with regular heating or microwave one    -   potassium tertiary-butoxide e.g. in DMSO,    -   sodium hydride e.g. in N,N-dimethylformamide with a regular        heating e.g. at reflux or with a microwave irradiation.        Optionally, before addition of the nitro derivative (VIII), the        phenol (IX) can be pre-stirred in presence of the solvent and        the base.

Compounds of formula (VIIc) wherein X═Y═N can be prepared bynucleophilic aromatic substitution from phenol (IX) and nitro compound(VIIIc) wherein usually Z═Cl. The base use is for example

-   -   potassium carbonate e.g. in N,N-dimethylformamide or        acetonitrile at room temperature    -   triethylamine e.g. in acetonitrile at reflux

Phenols of formula (IXa), wherein R¹ and R² are groups compatible withtypical nitrosation conditions, can be prepared using the correspondinganilines (X) with sodium nitrite in presence of an excess of acid suchas sulphuric acid in a solvent such as water, at 0° C. or 0° C.-5° C. ina first time and under heating in a second time e.g. at 40° C.-90° C.

Anilines of formula (Xa) (wherein R¹ and R² are groups non sensitive totypical nitrosation conditions) can be prepared from the nitroderivatives of formula (XI) (wherein R¹ and R² are groups non sensitiveto typical nitrosation conditions) using usual reduction conditions forexample

-   -   hydrogenation with H2 in a solvent, such as methanol in presence        of a metal catalyst such as Raney Nickel or Pd/C typically at        room temperature.    -   reduction in presence of Fe powder and ammonium chloride in a        solvent such as a mixture THF water or ethanol for example at        room temperature.

Some nitro derivatives of formula (XI) are commercially available.

Some other nitro derivatives such as the compound of formula (XIa)wherein R² is an ethoxy group can be prepared from the correspondingnitro-phenol derivative of formula (XII) by alkylation with for exampleethyl iodide in presence of a base such as potassium carbonate inacetone with heating e.g. at reflux.

Phenols of formula (IXb), wherein R¹ is H and R² is a C₁₋₄ alkoxy group(R⁵ is a C₁₋₄ alkyl group) can be prepared by monoalkylation from thecompounds of formula (XIII) using for example the suitable iodo-alkyl inpresence of a base such as potassium hydroxide in a solvent such asethanol.

step (iii): Compounds of formula (Ia), corresponding to compounds offormula (I) wherein X═C, Y═C can be prepared from compounds of formula(XIV) by heating e.g. at 100° C. in aqueous HCl.

step (ii): Compounds of formula (XIV) can be prepared from isocyanatesof formula (XV) wherein X═C, Y═C by addition of aminoacids of formula(VI) in presence of a base e.g. DIPEA in a solvent e.g. THF for exampleat room temperature.

step (i): Some isocyanates of formula (XV) are commercially availableother ones can be prepared from anilines of formula (IVb) usingtriphosgene and optionally triethylamine in a solvent such asdichloromethane at room temperature. Anilines (IVb) corresponding toanilines of formula (IV) with X═C, Y═C can be prepared with similarconditions to the ones described previously.

Optionally the two steps (ii) and (iii) can be performed in a one potfashion; in a first time, an addition of isocyanates (XV) overaminoacides of formula (VI) in presence of a base e.g. pyridine ortrietylamine in a solvent e.g. N,N-dimethylformamide or a mixturedichloromethane/DMF (e.g. at 35° C.), then in a second time addition ofHCl with heating (e.g. at 100° C.). Isocyanates can be prediluted in asolvent or not e.g. in THF.

Compounds of formula (Ib) corresponding to compounds of formula (I)wherein X═C, Y═C and R³═H can be prepared from anilines (IVb)[corresponding to anilines of formula (IV) wherein X═C, Y═C] in a onepot protocol.

In a first time, amino ester of formula (XVI) can be N-protected by aBoc group by reaction with Boc-anhydride in presence of a base e.g. DMAPin a solvent e.g. dichloromethane. In a second time, this solution canreact with anilines (IVb) with heating e.g. at 35° C. and in a thirdtime the cyclization can be promoted by addition of HCl and heating e.g.at 100° C.

step (ii): A nucleophilic substitution can be used to prepare compoundsof formula (Ic) corresponding to compounds of formula (I) wherein X═N,Y═C R³═R⁴=Me and (R¹, R²) are combinations of groups such as for example(p-CN, m-iPr), (H, m-OCF₃), (p-F, m-CH₃), (p-F, o-CH₃), (p-CN, m-Cl),(p-CN, o-Et), (p-Me, m-OCF₃), (p-CN, m-OMe). This reaction uses thecorresponding phenol of formula (IXc), a fluoro compound of formula(XVII) in presence of a base e.g. potassium carbonate heating e.g. at120° C. in a high boiling point solvent e.g. DMF.

step (i): Fluoro compound of formula (XVII) can be prepared byN-arylation of 5,5-dimethyl-2,4-imidazolidinedione (XVIII) with thearylboronic acid (XIX), promoted for example by copper (II) acetateusing a base e.g. pyridine and a solvent such as dichloromethane e.g. atroom temperature, open to air.

step (ii): Compounds of formula (Id) corresponding to compounds offormula (I) wherein X═N, Y═C or X═C, Y═C or X═N, Y═N and R³═R⁴=Me can beprepared from ureas of formula (XXa) by cyclization in presence of abase e.g. sodium methoxide heating e.g. at 65° C. in solvent such asmethanol.

step (i): Ureas of formula (XXa) can be prepared by addition of asolution containing an aniline of formula (IVc) (corresponding toanilines of formula IV wherein X═N, Y═C or X═C, Y═C or X═N, Y═N) and abase such as triethylamine in a solvent such as EtOAc to a solution of acarbonylating agent such as triphosgene in a solvent such as EtOAc e.g.at 0° C., followed by addition of triethylamine and ester (XXIa).Optionally, the ester (XXIa) can be pre-dissolved in a solvent such asEtOAc, the triethylamine being added to this pre-solution. Optionallysome additional triethylamine and ester (XXIa) or some additionaltriphosgene can be added. The needed ester (XXIa) can be prepared fromthe corresponding aminoacid, using methanol, heating the reactionmixture e.g. at reflux, after thionyl chloride addition.

step (ii): Compounds of formula (Ie) corresponding to compounds offormula (I) wherein X═N, Y═C or X═C, Y═C or X═N, Y═N and R³=Me, R⁴=Etcan be prepared from ureas of formula (XXb) by cyclization in presenceof a base e.g. sodium methoxide heating e.g. at 65° C. in solvent suchas methanol.

step (i): Ureas of formula (XXb) can be prepared by addition of asolution containing an aniline of formula (IVc) (corresponding toanilines of formula IV wherein X═N, Y═C or X═C, Y═C or X═N, Y═N) and abase such as triethylamine or diisopropylethylamine in a solvent such asEtOAc or dichloromethane to a solution of a carbonylating agent such astriphosgene in a solvent such as EtOAc or dichloromethane e.g. at 0° C.,followed by addition of triethylamine or diisopropylethylamine and ester(XXIb). Optionally, the ester (XXIb) can be pre-dissolved in a solventsuch as EtOAc or dichloromethane, the triethylamine ordiisopropylethylamine being added to this pre-solution. Optionally someadditional triethylamine or diisopropylethylamine and ester (XXIb) orsome additional triphosgene can be added. The needed ester (XXIb) can beprepared from the corresponding aminoacid, using methanol, heating thereaction mixture e.g. at reflux, after thionyl chloride addition.

step (iii): Phenols of formula (IX) can be prepared from compounds offormula (XXII) by removal of the benzyl group for example in presence ofhydrogen (e.g. P=1 atm) with a catalyst such as Pd/C in a solvent suchas methanol, a mixture of ethylacetate/ethanol etc.

step (ii): Compounds of formula (XXIIa) corresponding to compounds offormula (XXII) wherein [R¹; R²] are for example [(p-CN; o-Me or o-Et) or(p-Me or p-Et; m-OCF₃)] can be prepared by Negishi coupling using thesuitable pre-formed organozinc intermediate in solution (e.g. in THF)then Pd(tBu₃P)₂ and the corresponding bromo compound (XXIII) in asolvent such as THF. The organozinc intermediate can be prepared byaddition of a solution of zinc dichloride on the suitable alkylmagnesium bromide solution or by reverse addition of the alkyl magnesiumbromide solution on the zinc dichloride solution e.g. at −15° C., 0° C.or r.t. in solvents such as THF, diethyl ether. A solution of the bromocompound (XXIII) e.g. in THF can be added e.g. at 0° C. to theorganozinc intermediate or reversely the organozinc intermediatesolution can be added to a solution of the bromo compound (XXIII)pre-warmed e.g. at 60° C. Optionally some additional Pd(tBu₃P)₂ or someadditional pre-formed organozinc intermediate can be added.

step (i): Bromo compound of formula (XXIII) can be prepared fromcompounds of formula (XXIV) by benzylation with benzylhalide e.g. benzylbromide in presence of a base e.g. potassium carbonate, in a solventsuch as acetone heating e.g. at 50° C.

Compounds of formula (VIIc) [corresponding to compounds of formula (VII)wherein X═N, Y═C or X═N, Y═N, R¹═CN and R²=meta- or ortho- (Me, Et orcyclopropyl)] can be prepared by Negishi coupling using the suitablepre-formed organozinc intermediate in solution (e.g. in THF) thenPd(tBu₃P)₂ and the corresponding halo compounds (VIId) [corresponding tocompounds of formula (VII) wherein R¹═CN and R²=meta- or ortho-halogensuch as bromine and iodine)] in a solvent such as THF. The organozincintermediate can be prepared by addition of a solution of zincdichloride on the suitable alkyl magnesium bromide solution or byreverse addition of the alkyl magnesium bromide solution on the zincdichloride solution e.g. at −15° C., 0° C. or room temperature insolvents such as THF, diethyl ether. A solution of the halo compound(VIId) e.g. in THF can be added e.g. at 0° C. to the organozincintermediate or reversely the organozinc intermediate solution can beadded to a solution of the halo compound (VIId) pre-warmed e.g. at 60°C. Optionally some additional Pd(tBu₃P)₂ or some additional pre-formedorganozinc intermediate can be added.

Compounds of formula (XXV) wherein X═N, Y═C, R¹=para-CN andR⁵=meta-(cyclopropyl), meta-isopropenyl] can be prepared by Suzukicoupling using the corresponding boronic acid or boronic ester, a basesuch as potassium triphosphate, a system containing a palladium catalystand a ligand such as (Pd(OAc)₂/PCy₃) or (Pd(tBu₃)₂, in a solvent such asDMF, a mixture (toluene/water) etc heating e.g. at 110° C. optionallyunder microwave irradiation.

Phenol of formula (IXc) [corresponding to compound of formula (IX)wherein R¹=para-CN and R⁶=meta-I or ortho-Me] can be prepared fromfluoroaromatics of formula (XXVI), using potassium trimethylsilanolateand heating e.g. at a temperature ranging from r.t. to 70° C. in asolvent such as acetonitrile.

step ii: Phenol of formula (IXd) [corresponding to compound of formula(IX) wherein R¹=para-CN and R²=ortho-tBu] can be prepared from compoundsof formula (XXVII) using hydroxylamine hydrochloride in acetic acid,heating e.g. at reflux.

step i: Compound of formula (XXVII) can be prepared by a Reimer-Tiemannformylation starting from compound of formula (XXVIII) in a solvent suchas a mixture MeOH/water, using an hydroxide base such as sodiumhydroxide in water, heating e.g. at 60° C. and adding chloroform.

step ii: Compounds of formula (VIIf) [corresponding to compounds offormula (VII) wherein X═C, Y═N or X═C, Y═C and R¹=para-CN and R²═OR⁷with R⁷═C₁₋₄alkyl] can be prepared from compounds of formula (XXIX)using as alkylating agent a suitable halo derivative, a base such aspotassium carbonate in a solvent such as DMF e.g. at temperature rangingfrom r.t. to 60°

step i: Compounds of formula (XXIX) can be prepared from compounds offormula (XXX) and electrophiles (VIII), wherein Z═F or Cl in a similarmanner to the one described in scheme 5.

Phenol of formula (IXe) can be prepared from compound of formula (XXXI),using a demethylating agent such a BBr₃, in a solvent such asdichlorometane or dichloroethane at a suitable temperature ranging fromr.t. to 100° C. optionally under microwave irradiation.

step iii: Compounds of formula (IIIa) [corresponding to compounds offormula (III) wherein X═C, Y═N and R¹=para-CN and R²=iPr] can beprepared from compounds of formula (XXXII) by reduction with hydrogen(P=1 atm) in presence of a catalyst such as Pd/C, in a solvent such asmethanol.

step ii, i: Compounds of formula (XXXII) can be prepared in 2 steps fromnitro compounds of formula (XXXIII) using a similar way to the onedescribed on schemes 3, 2 (e.g. reduction with Fe/ammonium chloride andcoupling).

step i: Compounds of formula (XXXIV) wherein X═N, Y═N can be prepared bySuzuki coupling using the methyl boronic acid, a base such as potassiumtriphosphate, a system containing a palladium catalyst and a ligand suchas (Pd(OAc)₂/PCy₃) or (Pd(tBu₃)₂) in a solvent such as DMF heating e.g.at 110° C. optionally under microwave irradiation.

step i: Compounds of formula (Ie) can be prepared from compounds offormula (XXXV) by reduction with hydrogen (P=1 atm) in presence of acatalyst such as Pd/C, in a solvent such as methanol.

The present invention provides compounds of formula (I) or apharmaceutically acceptable salt thereof for use in therapy.

The compounds of formula (I) or their pharmaceutically acceptable saltsmay be of use for the treatment or prophylaxis of a disease or disorderwhere a modulator of the Kv3.1 or Kv3.2 or Kv3.1 and Kv3.2 channels isrequired. As used herein, a modulator of Kv3.1 or Kv3.2 or Kv 3.1 andKv3.2 is a compound which alters the properties of these channels,either positively or negatively.

Diseases or conditions that may be mediated by modulation of Kv3.1and/or Kv3.1 channels may be selected from the list below. The numbersin brackets after the listed diseases below refer to the classificationcode in Diagnostic and Statistical Manual of Mental Disorders, 4thEdition, published by the American Psychiatric Association (DSM-IV)and/or the International Classification of Diseases, 10th Edition(ICD-10).

The compounds of formula (I) or their pharmaceutically acceptable saltsmay be of use for the treatment or prophylaxis of depression and mooddisorders including Major Depressive Episode, Manic Episode, MixedEpisode and Hypomanic Episode; Depressive Disorders including MajorDepressive Disorder, Dysthymic Disorder (300.4), Depressive Disorder NotOtherwise Specified (311); Bipolar Disorders including Bipolar IDisorder, Bipolar II Disorder (Recurrent Major Depressive Episodes withHypomanic Episodes) (296.89), Cyclothymic Disorder (301.13) and BipolarDisorder Not Otherwise Specified (296.80); Other Mood Disordersincluding Mood Disorder Due to a General Medical Condition (293.83)which includes the subtypes With Depressive Features, With MajorDepressive-like Episode, With Manic Features and With Mixed Features),Substance-Induced Mood Disorder (including the subtypes With DepressiveFeatures, With Manic Features and With Mixed Features) and Mood DisorderNot Otherwise Specified (296.90);

Seasonal Affective Disorder.

The compounds of formula (I) or their pharmaceutically acceptable saltsmay be of use for the treatment or prophylaxis of schizophreniaincluding the subtypes Paranoid Type (295.30), Disorganised Type(295.10), Catatonic Type (295.20), Undifferentiated Type (295.90) andResidual Type (295.60); Schizophreniform Disorder (295.40);Schizoaffective Disorder (295.70) including the subtypes Bipolar Typeand Depressive Type; Delusional Disorder (297.1) including the subtypesErotomanic Type, Grandiose Type, Jealous Type, Persecutory Type, SomaticType, Mixed Type and Unspecified Type; Brief Psychotic Disorder (298.8);Shared Psychotic Disorder (297.3); Psychotic Disorder Due to a GeneralMedical Condition including the subtypes With Delusions and WithHallucinations; Substance-Induced Psychotic Disorder including thesubtypes With Delusions (293.81) and With Hallucinations (293.82); andPsychotic Disorder Not Otherwise Specified (298.9).

The compounds of formula (I) or their pharmaceutically acceptable saltsmay be of use for the treatment or prophylaxis of anxiety disordersincluding Panic Attack; Panic Disorder including Panic Disorder withoutAgoraphobia (300.01) and Panic Disorder with Agoraphobia (300.21);Agoraphobia; Agoraphobia Without History of Panic Disorder (300.22),Specific Phobia (300.29, formerly Simple Phobia) including the subtypesAnimal Type, Natural Environment Type, Blood-Injection-Injury Type,Situational Type and Other Type), Social Phobia (Social AnxietyDisorder, 300.23), Obsessive-Compulsive Disorder (300.3), PosttraumaticStress Disorder (309.81), Acute Stress Disorder (308.3), GeneralizedAnxiety Disorder (300.02), Anxiety Disorder Due to a General MedicalCondition (293.84), Substance-Induced Anxiety Disorder, SeparationAnxiety Disorder (309.21), Adjustment Disorders with Anxiety (309.24)and Anxiety Disorder Not Otherwise Specified (300.00):

The compounds of formula (I) or their pharmaceutically acceptable saltsmay be of use for the treatment or prophylaxis of substance-relateddisorders including Substance Use Disorders such as SubstanceDependence, Substance Craving and Substance Abuse; Substance-InducedDisorders such as Substance Intoxication, Substance Withdrawal,Substance-Induced Delirium, Substance-Induced Persisting Dementia,Substance-Induced Persisting Amnestic Disorder, Substance-InducedPsychotic Disorder, Substance-Induced Mood Disorder, Substance-InducedAnxiety Disorder, Substance-Induced Sexual Dysfunction,Substance-Induced Sleep Disorder and Hallucinogen Persisting PerceptionDisorder (Flashbacks); Alcohol-Related Disorders such as AlcoholDependence (303.90), Alcohol Abuse (305.00), Alcohol Intoxication(303.00), Alcohol Withdrawal (291.81), Alcohol Intoxication Delirium,Alcohol Withdrawal Delirium, Alcohol-Induced Persisting Dementia,Alcohol-Induced Persisting Amnestic Disorder, Alcohol-Induced PsychoticDisorder, Alcohol-Induced Mood Disorder, Alcohol-Induced AnxietyDisorder, Alcohol-Induced Sexual Dysfunction, Alcohol-Induced SleepDisorder and Alcohol-Related Disorder Not Otherwise Specified (291.9);Amphetamine (or Amphetamine-Like)-Related Disorders such as AmphetamineDependence (304.40), Amphetamine Abuse (305.70), AmphetamineIntoxication (292.89), Amphetamine Withdrawal (292.0), AmphetamineIntoxication Delirium, Amphetamine Induced Psychotic Disorder,Amphetamine-Induced Mood Disorder, Amphetamine-Induced Anxiety Disorder,Amphetamine-Induced Sexual Dysfunction, Amphetamine-Induced SleepDisorder and Amphetamine-Related Disorder Not Otherwise Specified(292.9); Caffeine Related Disorders such as Caffeine Intoxication(305.90), Caffeine-Induced Anxiety Disorder, Caffeine-Induced SleepDisorder and Caffeine-Related Disorder Not Otherwise Specified (292.9);Cannabis-Related Disorders such as Cannabis Dependence (304.30),Cannabis Abuse (305.20), Cannabis Intoxication (292.89), CannabisIntoxication Delirium, Cannabis-Induced Psychotic Disorder,Cannabis-Induced Anxiety Disorder and Cannabis-Related Disorder NotOtherwise Specified (292.9); Cocaine-Related Disorders such as CocaineDependence (304.20), Cocaine Abuse (305.60), Cocaine Intoxication(292.89), Cocaine Withdrawal (292.0), Cocaine Intoxication Delirium,Cocaine-Induced Psychotic Disorder, Cocaine-Induced Mood Disorder,Cocaine-Induced Anxiety Disorder, Cocaine-Induced Sexual Dysfunction,Cocaine-Induced Sleep Disorder and Cocaine-Related Disorder NotOtherwise Specified (292.9); Hallucinogen-Related Disorders such asHallucinogen Dependence (304.50), Hallucinogen Abuse (305.30),Hallucinogen Intoxication (292.89), Hallucinogen Persisting PerceptionDisorder (Flashbacks) (292.89), Hallucinogen Intoxication Delirium,Hallucinogen-Induced Psychotic Disorder, Hallucinogen-Induced MoodDisorder, Hallucinogen-Induced Anxiety Disorder and Hallucinogen-RelatedDisorder Not Otherwise Specified (292.9); Inhalant-Related Disorderssuch as Inhalant Dependence (304.60), Inhalant Abuse (305.90), InhalantIntoxication (292.89), Inhalant Intoxication Delirium, Inhalant-InducedPersisting Dementia, Inhalant-Induced Psychotic Disorder,Inhalant-Induced Mood Disorder, Inhalant-Induced Anxiety Disorder andInhalant-Related Disorder Not Otherwise Specified (292.9);Nicotine-Related Disorders such as Nicotine Dependence (305.1), NicotineWithdrawal (292.0) and Nicotine-Related Disorder Not Otherwise Specified(292.9); Opioid-Related Disorders such as Opioid Dependence (304.00),Opioid Abuse (305.50), Opioid Intoxication (292.89), Opioid Withdrawal(292.0), Opioid Intoxication Delirium, Opioid-Induced PsychoticDisorder, Opioid-Induced Mood Disorder, Opioid-Induced SexualDysfunction, Opioid-Induced Sleep Disorder and Opioid-Related DisorderNot Otherwise Specified (292.9); Phencyclidine (orPhencyclidine-Like)-Related Disorders such as Phencyclidine Dependence(304.60), Phencyclidine Abuse (305.90), Phencyclidine Intoxication(292.89), Phencyclidine Intoxication Delirium, Phencyclidine-InducedPsychotic Disorder, Phencyclidine-Induced Mood Disorder,Phencyclidine-Induced Anxiety Disorder and Phencyclidine-RelatedDisorder Not Otherwise Specified (292.9); Sedative-, Hypnotic-, orAnxiolytic-Related Disorders such as Sedative, Hypnotic, or AnxiolyticDependence (304.10), Sedative, Hypnotic, or Anxiolytic Abuse (305.40),Sedative, Hypnotic, or Anxiolytic Intoxication (292.89), Sedative,Hypnotic, or Anxiolytic Withdrawal (292.0), Sedative, Hypnotic, orAnxiolytic Intoxication Delirium, Sedative, Hypnotic, or AnxiolyticWithdrawal Delirium, Sedative-, Hypnotic-, or Anxiolytic-PersistingDementia, Sedative-, Hypnotic-, or Anxiolytic-Persisting AmnesticDisorder, Sedative-, Hypnotic-, or Anxiolytic-Induced PsychoticDisorder, Sedative-, Hypnotic-, or Anxiolytic-Induced Mood Disorder,Sedative-, Hypnotic-, or Anxiolytic-Induced Anxiety Disorder Sedative-,Hypnotic-, or Anxiolytic-Induced Sexual Dysfunction, Sedative-,Hypnotic-, or Anxiolytic-Induced Sleep Disorder and Sedative-,Hypnotic-, or Anxiolytic-Related Disorder Not Otherwise Specified(292.9); Polysubstance-Related Disorder such as Polysubstance Dependence(304.80); and Other (or Unknown) Substance-Related Disorders such asAnabolic Steroids, Nitrate Inhalants and Nitrous Oxide:

The compounds of formula (I) or their pharmaceutically acceptable saltsmay be of use for the treatment or prophylaxis of enhancement ofcognition including the treatment of cognition impairment in otherdiseases such as schizophrenia, bipolar disorder, depression, otherpsychiatric disorders and psychotic conditions associated with cognitiveimpairment, e.g. Alzheimer's disease.

The compounds of formula (I) or their pharmaceutically acceptable saltsmay be of use for the treatment or prophylaxis of sleep disordersincluding primary sleep disorders such as Dyssomnias such as PrimaryInsomnia (307.42), Primary Hypersomnia (307.44), Narcolepsy (347),Breathing-Related Sleep Disorders (780.59), Circadian Rhythm SleepDisorder (307.45) and Dyssomnia Not Otherwise Specified (307.47);primary sleep disorders such as Parasomnias such as Nightmare Disorder(307.47), Sleep Terror Disorder (307.46), Sleepwalking Disorder (307.46)and Parasomnia Not Otherwise Specified (307.47); Sleep Disorders Relatedto Another Mental Disorder such as Insomnia Related to Another MentalDisorder (307.42) and Hypersomnia Related to Another Mental Disorder(307.44); Sleep Disorder Due to a General Medical Condition, inparticular sleep disturbances associated with such diseases asneurological disorders, neuropathic pain, restless leg syndrome, heartand lung diseases; and Substance-Induced Sleep Disorder including thesubtypes Insomnia Type, Hypersomnia Type, Parasomnia Type and MixedType; sleep apnea and jet-lag syndrome:

The compounds of formula (I) or their pharmaceutically acceptable saltsmay be of use for the treatment or prophylaxis of eating disorders suchas Anorexia Nervosa (307.1) including the subtypes Restricting Type andBinge-Eating/Purging Type; Bulimia Nervosa (307.51) including thesubtypes Purging Type and Nonpurging Type; Obesity; Compulsive EatingDisorder; Binge Eating Disorder; and Eating Disorder Not OtherwiseSpecified (307.50):

The compounds of formula (I) or their pharmaceutically acceptable saltsmay be of use for the treatment or prophylaxis of Autism SpectrumDisorders including Autistic Disorder (299.00), Asperger's Disorder(299.80), Rett's Disorder (299.80), Childhood Disintegrative Disorder(299.10) and Pervasive Disorder Not Otherwise Specified (299.80,including Atypical Autism).

The compounds of formula (I) or their pharmaceutically acceptable saltsmay be of use for the treatment or prophylaxis ofAttention-Deficit/Hyperactivity Disorder including the subtypesAttention-Deficit/Hyperactivity Disorder Combined Type (314.01),Attention-Deficit/Hyperactivity Disorder Predominantly Inattentive Type(314.00), Attention-Deficit/Hyperactivity Disorder Hyperactive-ImpulseType (314.01) and Attention-Deficit/Hyperactivity Disorder Not OtherwiseSpecified (314.9); Hyperkinetic Disorder; Disruptive Behaviour Disorderssuch as Conduct Disorder including the subtypes childhood-onset type(321.81), Adolescent-Onset Type (312.82) and Unspecified Onset (312.89),Oppositional Defiant Disorder (313.81) and Disruptive Behaviour DisorderNot Otherwise Specified; and Tic Disorders such as Tourette's Disorder(307.23):

The compounds of formula (I) or their pharmaceutically acceptable saltsmay be of use for the treatment or prophylaxis of Personality Disordersincluding the subtypes Paranoid Personality Disorder (301.0), SchizoidPersonality Disorder (301.20), Schizotypal Personality Disorder(301,22), Antisocial Personality Disorder (301.7), BorderlinePersonality Disorder (301,83), Histrionic Personality Disorder (301.50),Narcissistic Personality Disorder (301,81), Avoidant PersonalityDisorder (301.82), Dependent Personality Disorder (301.6),Obsessive-Compulsive Personality Disorder (301.4) and PersonalityDisorder Not Otherwise Specified (301.9).

The compounds of formula (I) or their pharmaceutically acceptable saltsmay be of use for the treatment or prophylaxis of Sexual dysfunctionsincluding Sexual Desire Disorders such as Hypoactive Sexual DesireDisorder (302.71), and Sexual Aversion Disorder (302.79); sexual arousaldisorders such as Female Sexual Arousal Disorder (302.72) and MaleErectile Disorder (302.72); orgasmic disorders such as Female OrgasmicDisorder (302.73), Male Orgasmic Disorder (302.74) and PrematureEjaculation (302.75); sexual pain disorder such as Dyspareunia (302.76)and Vaginismus (306.51); Sexual Dysfunction Not Otherwise Specified(302.70); paraphilias such as Exhibitionism (302.4), Fetishism (302.81),Frotteurism (302.89), Pedophilia (302.2), Sexual Masochism (302.83),Sexual Sadism (302.84), Transvestic Fetishism (302.3), Voyeurism(302.82) and Paraphilia Not Otherwise Specified (302.9); gender identitydisorders such as Gender Identity Disorder in Children (302.6) andGender Identity Disorder in Adolescents or Adults (302.85); and SexualDisorder Not Otherwise Specified (302.9).

The compounds of formula (I) or their pharmaceutically acceptable saltsmay be of use for the treatment or prophylaxis of Impulse controldisorder” including: Intermittent Explosive Disorder (312.34),Kleptomania (312.32), Pathological Gambling (312.31), Pyromania(312.33), Trichotillomania (312.39), Impulse-Control Disorders NotOtherwise Specified (312.3), Binge Eating, Compulsive Buying, CompulsiveSexual Behaviour and Compulsive Hoarding.

The compounds of formula (I) or their pharmaceutically acceptable saltsmay be of use for the treatment or prophylaxis of hearing disordersincluding auditory neuropathy, auditory processing disorder, hearingloss, which includes sudden hearing loss, noise induced hearing loss,substance-induced hearing loss, and hearing loss in adults over 60(presbycusis), and tinnitus.

The compounds of formula (I) or their pharmaceutically acceptable saltsmay be of use for the treatment or prophylaxis of Ménière's disease,disorders of balance, and disorders of the inner ear.

The compounds of formula (I) or their pharmaceutically acceptable saltsmay be of use for the treatment or prophylaxis of hyperacusis anddisturbances of loudness perception, including Fragile-X syndrome andautism.

The compounds of formula (I) or their pharmaceutically acceptable saltsmay be of use for the treatment or prophylaxis of Epilepsy, (including,but not limited to, localization-related epilepsies, generalizedepilepsies, epilepsies with both generalized and local seizures, and thelike), seizures associated with Lennox-Gastaut syndrome, seizures as acomplication of a disease or condition (such as seizures associated withencephalopathy, phenylketonuria, juvenile Gaucher's disease, Lundborg'sprogressive myoclonic epilepsy, stroke, head trauma, stress, hormonalchanges, drug use or withdrawal, alcohol use or withdrawal, sleepdeprivation, fever, infection, and the like), essential tremor, restlesslimb syndrome, partial and generalised seizures (including tonic,clonic, tonic-clonic, atonic, myoclonic, absence seizures), secondarilygeneralized seizures, temporal lobe epilepsy, absence epilepsies(including childhood, juvenile, myoclonic, photo- and pattern-induced),severe epileptic encephalopathies (including hypoxia-related andRasmussen's syndrome), febrile convulsions, epilepsy partialis continua,progressive myoclonus epilepsies (including Unverricht-Lundborg diseaseand Lafora's disease), post-traumatic seizures/epilepsy including thoserelated to head injury, simple reflex epilepsies (includingphotosensive, somatosensory and proprioceptive, audiogenic andvestibular), metabolic disorders commonly associated with epilepsy suchas pyridoxine-dependent epilepsy, Menkes' kinky hair disease, Krabbe'sdisease, epilepsy due to alcohol and drug abuse (e.g. cocaine), corticalmalformations associated with epilepsy (e.g. double cortex syndrome orsubcortical band heterotopia), chromosomal anomolies associated withseizures or epilepsy such as Partial monosomy (15Q)/Angelman syndrome)

and the like.

In one embodiment of the invention, there is provided a compound offormula (I) or a pharmaceutically acceptable salt thereof for thetreatment or prophylaxis of depression and mood disorders, hearingdisorders, schizophrenia, substance abuse disorders, sleep disorders orepilepsy.

In one embodiment of the invention, there is provided a compound offormula (I) or a pharmaceutically acceptable salt thereof for thetreatment or prophylaxis of bipolar disorder or mania.

The term “treatment” or “treating” as used herein includes the control,mitigation, reduction, or modulation of the disease state or itssymptoms.

The term “prophylaxis” is used herein to mean preventing symptoms of adisease or disorder in a subject or preventing recurrence of symptoms ofa disease or disorder in an afflicted subject and is not limited tocomplete prevention of an affliction.

The invention also provides a method of treating or preventing a diseaseor disorder where a modulator of Kv3 is required, for example thosediseases and disorders mentioned hereinabove, which comprisesadministering to a subject in need thereof an effective amount of acompound of formula (I) or a pharmaceutically acceptable salt thereof.

The invention also provides a compound of formula (I), or apharmaceutically acceptable salt thereof, for use in the treatment orprophylaxis of a disease or disorder where a modulator of Kv3 isrequired, for example those diseases and disorders mentionedhereinabove.

The invention also provides the use of a compound of formula (I), or apharmaceutically acceptable salt thereof, in the manufacture of amedicament for the treatment or prophylaxis of a disease or disorderwhere a modulator of Kv3 is required, for example those diseases anddisorders mentioned hereinabove.

The invention also provides a method of treating depression and mooddisorders, schizophrenia, substance abuse disorders, sleep disorders orepilepsy, for example for those indications mentioned hereinabove, whichcomprises administering to a subject in need thereof an effective amountof a Kv3 modulator or a pharmaceutically acceptable salt thereof.

For use in therapy the compounds of the invention are usuallyadministered as a pharmaceutical composition. The invention alsoprovides a pharmaceutical composition comprising a compound of formula(I), or a pharmaceutically acceptable salt thereof, and apharmaceutically acceptable carrier.

The compounds of formula (I) or their pharmaceutically acceptable saltsmay be administered by any convenient method, e.g. by oral, parenteral,buccal, sublingual, nasal, rectal or transdermal administration, and thepharmaceutical compositions adapted accordingly.

The compounds of formula (I) or their pharmaceutically acceptable saltswhich are active when given orally can be formulated as liquids orsolids, e.g. as syrups, suspensions, emulsions, tablets, capsules orlozenges.

A liquid formulation will generally consist of a suspension or solutionof the active ingredient in a suitable liquid carrier(s) e.g. an aqueoussolvent such as water, ethanol or glycerine, or a non-aqueous solvent,such as polyethylene glycol or an oil. The formulation may also containa suspending agent, preservative, flavouring and/or colouring agent.

A composition in the form of a tablet can be prepared using any suitablepharmaceutical carrier(s) routinely used for preparing solidformulations, such as magnesium stearate, starch, lactose, sucrose andcellulose.

A composition in the form of a capsule can be prepared using routineencapsulation procedures, e.g. pellets containing the active ingredientcan be prepared using standard carriers and then filled into a hardgelatin capsule; alternatively a dispersion or suspension can beprepared using any suitable pharmaceutical carrier(s), e.g. aqueousgums, celluloses, silicates or oils and the dispersion or suspensionthen filled into a soft gelatin capsule.

Typical parenteral compositions consist of a solution or suspension ofthe active ingredient in a sterile aqueous carrier or parenterallyacceptable oil, e.g. polyethylene glycol, polyvinyl pyrrolidone,lecithin, arachis oil or sesame oil. Alternatively, the solution can belyophilised and then reconstituted with a suitable solvent just prior toadministration.

Compositions for nasal administration may conveniently be formulated asaerosols, drops, gels and powders. Aerosol formulations typicallycomprise a solution or fine suspension of the active ingredient in apharmaceutically acceptable aqueous or non-aqueous solvent and areusually presented in single or multidose quantities in sterile form in asealed container which can take the form of a cartridge or refill foruse with an atomizing device. Alternatively the sealed container may bea disposable dispensing device such as a single dose nasal inhaler or anaerosol dispenser fitted with a metering valve. Where the dosage formcomprises an aerosol dispenser, it will contain a propellant which canbe a compressed gas e.g. air, or an organic propellant such as afluorochloro-hydrocarbon or hydrofluorocarbon. Aerosol dosage forms canalso take the form of pump-atomizers.

Compositions suitable for buccal or sublingual administration includetablets, lozenges and pastilles where the active ingredient isformulated with a carrier such as sugar and acacia, tragacanth, orgelatin and glycerin.

Compositions for rectal administration are conveniently in the form ofsuppositories containing a conventional suppository base such as cocoabutter.

Compositions suitable for transdermal administration include ointments,gels and patches.

In one embodiment the composition is in unit dose form such as a tablet,capsule or ampoule.

The composition may contain from 0.1% to 100% by weight, for examplefrom 10 to 60% by weight, of the active material, depending on themethod of administration. The composition may contain from 0% to 99% byweight, for example 40% to 90% by weight, of the carrier, depending onthe method of administration. The composition may contain from 0.05 mgto 1000 mg, for example from 1.0 mg to 500 mg, of the active material,depending on the method of administration. The composition may containfrom 50 mg to 1000 mg, for example from 100 mg to 400 mg of the carrier,depending on the method of administration. The dose of the compound usedin the treatment of the aforementioned disorders will vary in the usualway with the seriousness of the disorders, the weight of the sufferer,and other similar factors. However, as a general guide suitable unitdoses may be 0.05 to 1000 mg, more suitably 1.0 to 500 mg, and such unitdoses may be administered more than once a day, for example two or threea day. Such therapy may extend for a number of weeks or months.

The invention provides, in a further aspect, a combination comprising acompound of formula (I) or a pharmaceutically acceptable derivativethereof together with a further therapeutic agent or agents.

When the compounds are used in combination with other therapeuticagents, the compounds may be administered either sequentially orsimultaneously by any convenient route.

The combinations referred to above may conveniently be presented for usein the form of a pharmaceutical formulation and thus pharmaceuticalformulations comprising a combination as defined above together with apharmaceutically acceptable carrier or excipient comprise a furtheraspect of the invention. The individual components of such combinationsmay be administered either sequentially or simultaneously in separate orcombined pharmaceutical formulations.

When a compound of formula (I) or a pharmaceutically acceptablederivative thereof is used in combination with a second therapeuticagent active against the same disease state the dose of each compoundmay differ from that when the compound is used alone. Appropriate doseswill be readily appreciated by those skilled in the art.

A pharmaceutical composition of the invention, which may be prepared byadmixture, suitably at ambient temperature and atmospheric pressure, isusually adapted for oral, parenteral or rectal administration and, assuch, may be in the form of tablets, capsules, oral liquid preparations,powders, granules, lozenges, reconstitutable powders, injectable orinfusible solutions or suspensions or suppositories. Orallyadministrable compositions are generally preferred.

The present invention also provides Kv3 modulators, or theirpharmaceutically acceptable salts, for use in the treatment orprophylaxis of depression and mood disorders, hearing disorders,schizophrenia, substance abuse disorders, sleep disorders or epilepsy.

In particular Kv3 modulators or their pharmaceutically acceptable saltsmay be particularly useful in the treatment or prophylaxis of depressionand mood disorders including Major Depressive Episode, Manic Episode,Mixed Episode and Hypomanic Episode; Depressive Disorders includingMajor Depressive Disorder, Dysthymic Disorder (300.4), DepressiveDisorder Not Otherwise Specified (311); Bipolar Disorders includingBipolar I Disorder, Bipolar II Disorder (Recurrent Major DepressiveEpisodes with Hypomanic Episodes) (296.89), Cyclothymic Disorder(301.13) and Bipolar Disorder Not Otherwise Specified (296.80); OtherMood Disorders including Mood Disorder Due to a General MedicalCondition (293.83) which includes the subtypes With Depressive Features,With Major Depressive-like Episode, With Manic Features and With MixedFeatures), Substance-Induced Mood Disorder (including the subtypes WithDepressive Features, With Manic Features and With Mixed Features) andMood Disorder Not Otherwise Specified (296.90), Seasonal affectivedisorder

The invention also provides a method of treating depression and mooddisorders, hearing disorders, schizophrenia, substance abuse disorders,sleep disorders or epilepsy, including for example those disordersmentioned hereinabove, which comprises administering to a subject inneed thereof an effective amount of Kv3 modulator or a pharmaceuticallyacceptable salt thereof.

The invention also provides a Kv3 modulator, or a pharmaceuticallyacceptable salt thereof, for use in the treatment or prophylaxis ofdepression and mood disorders, hearing disorders, schizophrenia,substance abuse disorders, sleep disorders or epilepsy, including forexample those disorders mentioned hereinabove.

The invention also provides the use of a Kv3 modulator, or apharmaceutically acceptable salt thereof, in the manufacture of amedicament for the treatment or prophylaxis of depression and mooddisorders, hearing disorders, schizophrenia, substance abuse disorders,sleep disorders or epilepsy, including for example those disordersmentioned hereinabove.

For use in therapy the Kv3 modulators are usually administered as apharmaceutical composition for example a composition comprising a Kv3modulator or a pharmaceutically acceptable salt thereof, and apharmaceutically acceptable carrier. Examples of such compositions, andmethods of administration thereof, which compositions comprise acompound of formula (I) or a pharmaceutically acceptable salt thereof,are described hereinabove. Such compositions and methods ofadministration may also be used for other Kv3 modulators orpharmaceutically acceptable salts thereof, in the treatment ofdepression and mood disorders, hearing disorders, schizophrenia,substance abuse disorders, sleep disorders or epilepsy, including forexample those disorders mentioned hereinabove.

All publications, including but not limited to patents and patentapplications, cited in this specification are herein incorporated byreference as if each individual publication were specifically andindividually indicated to be incorporated by reference herein as thoughfully set forth.

BRIEF DESCRIPTION OF THE DRAWINGS

The present invention is illustrated, by way of example only, withreference to the following figures in which:

FIG. 1 a hKv3.2 currents recorded using the assay described in Example89. Data shown are the individual currents over the period of thedepolarizing voltage step to −15 mV recorded from 4 different cells attwo concentrations of the compound of Example 19. The data are fitted bya single exponential curve (solid lines) using the fitting procedure inPrism version 5 (Graphpad Software Inc).

FIG. 1 b hKv3.2 currents recorded using the assay described in Example89. Data shown are the individual currents over the period of thedepolarizing voltage step to −15 mV recorded from 2 different cells attwo concentrations of compound of Example 71. The data are fitted by asingle exponential curve (solid lines) using the fitting procedure inPrism version 5 (Graphpad Software Inc).

FIG. 2 Recordings made from identified “fast-firing” interneurons in thesomatosensory cortex of the mouse.

FIG. 3 The frequency of action potentials recorded fromparvalbumin-positive interneurons in the somatosensory cortex of themouse, evoked by depolarizing current steps

FIG. 4 The half-width of evoked action potentials fromparvalbumin-positive interneurons in the somatosensory cortex of themouse FIG. 5 High-voltage activated potassium currents recorded fromvisually identified MNTB neurons in the mouse, in vitro

EXPERIMENTAL

The invention is illustrated by the Compounds described below. In theprocedures that follow, after each starting material, reference to adescription is typically provided. This is provided merely forassistance to the skilled chemist. The starting material may notnecessarily have been prepared from the Description referred to.

Analytical Equipment

Starting materials, reagents and solvents were obtained from commercialsuppliers and used without further purification unless otherwise stated.Unless otherwise stated, all compounds with chiral centres are racemic.Where reactions are described as having been carried out in a similarmanner to earlier, more completely described reactions, the generalreaction conditions used were essentially the same. Work up conditionsused were of the types standard in the art, but may have been adaptedfrom one reaction to another. The starting material may not necessarilyhave been prepared from the batch referred to Compounds synthesised mayhave various purities ranging from for example 85% to 98%.

Proton Magnetic Resonance (NMR) spectra were recorded either on Varianinstruments at 300, 400, 500 or 600 MHz, or on Bruker instruments at 400MHz. Chemical shifts are reported in ppm (δ) using the residual solventline as internal standard. Splitting patterns are designed as s(singlet), br.s (broad singlet), d (doublet), t (triplet), q (quartet),dd (doublet of doublets), dt (doublet of triplets) and m (multiplet).The NMR spectra were recorded at temperatures ranging from 25 to 30° C.

Direct infusion Mass spectra (MS) were run on an Agilent 1100 SeriesLC/MSD Mass Spectrometer, operating in ES (+) and ES (−) ionization mode[ES (+): Mass range: 100-1000 amu. Infusion solvent: water+0.1%HCO2H/CH3CN 50/50. ES (−): Mass range: 100-1000 amu. Infusion solvent:water+0.05% NH4OH/CH3CN 50/50]. The use of this methodology is indicatedby “MS_(—)1 (ESI)” in the analytic characterization of the describedcompounds. Alternatively, Mass spectra (MS) were run on a massspectrometer, operating in ES (+) and ES (−) ionization mode coupledwith an HPLC instrument Agilent 1100 Series [LC/MS-ESI(+) analyses wereperformed on a Supelcosil ABZ+Plus (33×4.6 mm, 3 μm) (mobile phase: from10%[CH₃CN+0.05% TFA] to 90%[CH₃CN+0.05% TFA] and 10% [water] in 2.2 min,under these conditions for 2.8 min. T=45° C., flux=0.9 mL/min)]. The useof this methodology is indicated by “MS_(—)2 (ESI)” in the analyticcharacterization of the described compounds.

HPLC-Mass spectra (HPLC-MS) were taken on an Agilent 1100 Series LC/MSDMass Spectrometer coupled with HPLC instrument Agilent 1100 Series,operating in positive or negative electrospray ionization mode and inboth acidic and basic gradient conditions. Acidic gradient: LC/MS-ES (+or −) analyses were performed on a Supelcosil ABZ+Plus column (33×4.6mm, 3 μm). Mobile phase: A: (water+0.1% HCO2H)/B: CH3CN. Gradient(standard method): t=0 min 0% (B), from 0% (B) to 95% (B) in 5 minlasting for 1.5 min, from 95% (B) to 0% (B) in 0.1 min, stop time 8.5min. Column T=r.t. Flow rate=1 ml/min. The use of this methodology isindicated by “LC-MS_A” in the analytic characterization of the describedcompounds.

Ultra Performance Liquid Chromatography with an Acidic Gradient:

Total ion current (TIC) and DAD UV chromatographic traces together withMS and UV spectra associated with the peaks were taken on a UPLC/MSAcquity™ system equipped with 2996 PDA detector and coupled to a WatersMicromass ZQ™ mass spectrometer operating in positive or negativeelectrospray ionisation mode [LC/MS−ES (+ or −): analyses were performedusing an Acquity™ UPLC BEH C18 column (50×2.1 mm, 1.7 μm particle size).

General Method:

Mobile phase: A: (water+0.1% HCO2H)/B: (CH3CN+0.06% HCO2H). Gradient:t=0 min 3% (B), t=0.05 min 6% (B), t=0.57 min 70% (B), t=1.06 min 99%(B) lasting for 0.389 min, t=1.45 min 3% (B), stop time 1.5 min. ColumnT=40° C. Flow rate=1.0 mL/min. Mass range: ES (+): 100-1000 amu. ES (−):100-800 amu. UV detection range: 210-350 nm. The use of this methodologyis indicated by “UPLC” in the analytic characterization of the describedcompounds.

1^(st) Focussed Method:

Mobile phase: A: (water+0.1% HCO2H)/B: (CH3CN+0.1% HCO2H). Gradient: t=0min 3% (B), t=1.06 min 99% (B), t=1.45 min 99% (B), t=1.46 min 3% (B),stop time 1.5 min. Column T=40° C. Flow rate=1.0 mL/min. Mass range: ES(+): 100-1000 amu. ES (−): 100-800 amu. UV detection range: 210-350 nm.The use of this methodology is indicated by “UPLC_s” in the analyticcharacterization of the described compounds.

2^(nd) Focussed Method:

Mobile phase: A: (water+0.1% HCO2H)/B: (CH3CN+0.1% HCO2H). Gradient: t=0min 3% (B), t=1.5 min 100% (B), t=1.9 min 100% (B), t=2 min 3% (B), stoptime 2 min. Column T=40° C. Flow rate=1.0 mL/min. Mass range: ES (+):100-1000 amu. ES (−): 100-800 amu. UV detection range: 210-350 nm. Theuse of this methodology is indicated by “UPLC_ipqc” in the analyticcharacterization of the described compounds.

Ultra Performance Liquid Chromatography with a Basic Gradient:

Total ion current (TIC) and DAD UV chromatographic traces together withMS and UV spectra associated with the peaks were taken on a UPLC/MSAcquity™ system equipped with PDA detector and coupled to a Waters SQDmass spectrometer operating in positive and negative alternateelectrospray ionisation mode [LC/MS−ES+/−: analyses were performed usingan Acquity™ UPLC BEH C18 column (50×2.1 mm, 1.7 μm particle size).Mobile phase: A: (10 mM aqueous solution of NH4HCO3 (adjusted to pH 10with ammonia))/B: CH3CN. Gradient: t=0 min 3% (B), t=1.06 min 99% (B)lasting for 0.39 min, t=1.46 min 3% (B), stop time 1.5 min. Column T=40°C. Flow rate=1.0 mL/min. Mass range: ES (+): 100-1000 amu. ES (−):100-1000 amu. UV detection range: 220-350 nm. The use of thismethodology is indicated by “UPLC_B” in the analytic characterization ofthe described compounds.

For reactions involving microwave irradiation, a Personal ChemistryEmrys™ Optimizer was used or a Biotage Initiator

In a number of preparations, purification was performed using Biotagemanual flash chromatography (Flash+), Biotage automatic flashchromatography (Horizon, SP1 and SP4), Companion CombiFlash (ISCO)automatic flash chromatography, Flash Master Personal or Vac Mastersystems.

Flash chromatographies were carried out on silica gel 230-400 mesh(supplied by Merck AG Darmstadt, Germany) or on silica gel 300-400 mesh(supplied by Sinopharm Chemical Reagent Co., Ltd.), Varian Mega Be-Sipre-packed cartridges, pre-packed Biotage silica cartridges (e.g.Biotage SNAP cartridge), KP-NH prepacked flash cartridges, ISOLUTE NH₂prepacked cartridges or ISCO RediSep Silica cartridges.

SPE-Si cartridges are silica solid phase extraction columns supplied byVarian.

In a number of preparations, purification was performed on aMass-Directed Autopurification (MDAP) system Fractionlynx™ equipped withWaters 2996 PDA detector and coupled with ZQ™ mass spectrometer (Waters)operating in positive and negative electrospray ionisation mode ES+, ES−(mass range 100-1000 or 100-900) A set of semi-preparative gradientshave been used:

Method A:

Chromatographic Basic Conditions

Column: XTerra Prep MS C18 OBD (150 mm×30 mm 10 μm particle size) at

room temperature

Mobile phase: A: (water+10 mM aqueous solution of ammonium bicarbonate(adjusted to

-   -   pH 10 with ammonia)), B: acetonitrile

Flow rate: 40 ml/min

Gradient: 10% (B) for 0.5 min, from 10% (B) to 95% (B) in 12.5 min, from95% (B) to 100% (B) in 3 min

Method B:

Chromatographic Basic Conditions

Column: XTerra Prep MS C18 OBD (150 mm×30 mm 10 μm particle size) at

room temperature

Mobile phase: A: water+10 mM aqueous solution of ammonium bicarbonate(adjusted to

-   -   pH 10 with ammonia), B: acetonitrile

Flow rate: 40 ml/min

Gradient: from 20% to 25% (B) in 1 min, from 25% (B) to 65% (B) in 12min, from 65% (B) to 100% (B) in 0.5 min

Method C:

Chromatographic Basic Conditions

Column: Waters Xbridge C18 OBD (50 mm×19 mm 5 μm particle size) at

room temperature

Mobile phase: A: water+10 mM aqueous solution of ammonium bicarbonate(adjusted to

-   -   pH 10 with ammonia), B: acetonitrile

Flow rate: 17 ml/min

Gradient: from 20% (B) to 25% (B) in 1 min, from 25% (B) to 55% (B) in 9min, from 55% (B) to 100% (B) in 2 min, return to 20% (B) in 0.1 min

Method D:

Chromatographic Acidic Conditions

Column: Waters Xbridge C18 OBD (50 mm×19 mm 5 μm particle size) at

room temperature

Mobile phase: A: (water+0.1% formic acid in water); B: acetonitrile

Flow rate: 17 ml/min

Gradient: from 20% (B) to 25% B in 1 min, from 25% (B) to 55% (B) in 9min, from 55% (B) to

-   -   100% (B) in 2 min, return to 20% (B) in 0.1 min

Method E:

Chromatographic Basic Conditions

Column: Waters Xbridge C18 OBD (50 mm×19 mm 5 μm particle size) at

room temperature

Mobile phase: A: (water+10 mM aqueous solution of ammonium bicarbonate(adjusted to

pH 10 with ammonia)), B: acetonitrile

Flow rate: 17 ml/min

Gradient: from 10% (B) to 15% (B) in 1 min, from 15% (B) to 70% (B) in 7min, from 70% (B)

-   -   to 100% (B) in 1 min, 100% (B) for 2 min, return to 10% (B) in        0.1 min

Method F:

Chromatographic Basic Conditions

Column: Phenomenex Gemini AXIA C18 (50×21.2 mm 5 μm particle size)

Mobile phase: A: water+10 mM aqueous solution of ammonium bicarbonate(adjusted to

pH 10 with ammonia), B: acetonitrile

Flow rate: 17 ml/min

Gradient: from 10% (B) to 15% (B) in 1 min, from 15% (B) to 65% (B) in 8min, from 65% (B) to 100% (B) in 1 min, return to 10% (B) in 1 min.

Method G:

Chromatographic Basic Conditions

Column: Phenomenex Gemini AXIA C18 (50×21.2 mm 5 μm particle size)

Mobile phase: A: water+10 mM aqueous solution of ammonium bicarbonate(adjusted to

pH 10 with ammonia), B: acetonitrile

Flow rate: 17 ml/min

Gradient: from 10% (B) to 15% (B) in 1 min, from 15% (B) to 70% (B) in 7min, from 70% (B) to 100% (B) in 1 min, 100% (B) during 2 min, return to10% (B) in 0.1 min.

Method H:

Chromatographic Acidic Conditions

Column: Waters Xbridge C18 OBD (100 mm×19 mm 5 μm particle size) at roomtemperature

Mobile phase: A: (water+0.1% formic acid in water); B: acetonitrile

Flow rate: 17 ml/min

Gradient: 5% (B) during 1 min, from 5% (B) to 90% (B) in 9 min, from 90%(B) to

-   -   100% (B) in 0.1 min, 100% (B) during 0.8 min, return to 5% (B)        in 0.1 min

Method I:

Chromatographic Acidic Conditions

Column: Waters Sunfire OBD (100 mm×19 mm, 5 μm particle size) at roomtemperature

Mobile phase: A: (water+0.1% formic acid in water); B: acetonitrile

Flow rate: 17 ml/min

Gradient: from 30% (B) to 70% (B) in 9 min, from 70% (B) to 100% (B) in1 min, return to 30% (B) then 30% (B) during 1 min

Method J:

Chromatographic Acidic Conditions

Column: Waters Sunfire OBD (100 mm×19 mm, 5 μm particle size) at roomtemperature

Mobile phase: A: (water+0.1% formic acid in water); B: acetonitrile

Flow rate: 17 mL/min

Gradient: 10% (B) during 1 min, from 10% (B) to 95% (B) in 10 min, 95%(B) during 1.5 min,

return to 10% (B) in 0.1 min.

SPE-SCX cartridges are ion exchange solid phase extraction columnssupplied by Varian. The eluent used with SPE-SCX cartridges is DCM andMeOH or MeCN or MeOH followed by ammonia solution in MeOH (typically 2N). The collected fractions are those eluted with the ammonia solutionin MeOH unless otherwise stated.

ABBREVIATIONS

Boc t-butyloxycarbonyl

CDCl₃ deutrated chloroform

CH3CN acetonitrile

(CH₂O)_(n) paraformaldehyde

cHex cyclohexane

CV column volume

(Cy)₃P Tricyclohexylphosphine

DCM dichloromethane

DIPEA N,N-diisopropylethylamine

DMAP 4-dimethylaminopyridine

DMF N,N-dimethylformamide

DMSO dimethylsulfoxide

DMSO-d₆ deutrated dimethylsulfoxide

EDC.HCl N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride

Et₂O diethyl ether

EtOAc ethyl acetate

h hours

H₂ gaseous hydrogen

HATU (O-7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate)

HBTU O-benzotriazol-1-yl-tetramethyluronium hexafluorophosphate

HCO2H formic acid

HCl hydrogen chloride

HNO₃ nitric acid

HOBt. H₂O 1-hydroxybenzyltriazole hydrate

H₂SO₄ sulfuric acid

K₂CO₃ potassium carbonate

KOH potassium hydroxide

MeCN/CH₃CN acetonitrile

MeOH methanol

methanol-d₄ deutrated methanol

MDAP mass-directed autopurification

N₂ gaseous nitrogen

NaBH(OAc)₃ sodium triacethoxyborohydride

NaHCO₃ sodium hydrogenocarbonate

NaNO₂ sodium nitrite

Na₂CO₃ sodium carbonate

NaOH sodium hydroxide

NH4OH ammonium hydroxide

NH4 HCO3H ammonium bicarbonate

NMR Nuclear Magnetic Resonance

Pd/C palladium on charcoal

Pd(OAc)₂ Palladium(II) acetate

Pd(tBu₃P)₂ Palladium (0) bis(Tri-Tert-Butylphosphine)

PE petroleum ether

r.t. room temperature

tBuOK potassium tert-butoxide

TBTU o-Benzotriazol-1-yl-n,n,n′,n′-tetramethyluronium tetrafluoroborate

TEA triethylamine

TFA trifluoroacetic acid

THF tetrahydrofuran

TsOH*H₂O 4-methylbenzenesulfonic acid hydrate, p-toluenesulfonic acidhydrate

Supporting Examples and Intermediates Intermediate 11,1-dimethylethyl{(1R)-methyl-2-[(4-{[3-(methyloxy)phenyl]oxy}phenyl)amino]-2-oxoethyl}carbamate

To a solution of N-{[(1,1-dimethylethyl)oxy]carbonyl}-D-alanine (250 mg,1.321 mmol) in dry N,N-dimethylformamide (5 mL), DIPEA (0.346 mL, 1.982mmol) and then TBTU (467 mg, 1.453 mmol) were added and the reactionmixture was stirred for 15 minutes at room temperature.(4-{[3-(methyloxy)phenyl]oxy}phenyl)amine (313 mg, 1.453 mmol) was thenadded and the reaction mixture was stirred for 30 minutes at roomtemperature. The reaction was quenched with brine (10 mL), diluted withwater (5 mL) and extracted with diethyl ether (3 times 20 mL). Theorganic layer was dried over sodium sulphate, filtered and evaporatedand the residue was purified by silica gel chromatography (Biotagesystem, 25 g, SNAP column) using as eluents a gradient cyclohexane/ethylacetate from 100/0 to 70/30 to afford the title compound (440 mg) as alight yellow gum.

¹H-NMR (400 MHz, DMSO-d₆) δ ppm: 9.96 (1H, s), 7.62 (2H, m), 7.25 (1H,t), 7.04-7.11 (1H, m), 7.01 (2H, m), 6.68 (1H, dd), 6.44-6.55 (2H, m),4.04-4.16 (1H, m), 3.70-3.76 (3H, m), 1.29-1.46 (9H, m), 1.19-1.29 (3H,m); UPLC: 0.76 min, 387 [M+H]+.

Intermediate 2 N¹-(4-{[3-(methyloxy)phenyl]oxy}phenyl)-D-alaninamide

To a solution of 1,1-dimethylethyl{(1R)-1-methyl-2-[(4-{[3-(methyloxy)phenyl]oxy}phenyl)amino]-2-oxoethyl}carbamate(Intermediate 1, 435 mg) in dry dichloromethane (6 mL), TFA (2 mL, 26.0mmol) was added and the reaction mixture was stirred for 1 hour at roomtemperature. The solvent and the excess of TFA were evaporated and theresidue was purified by SCX cartridge to afford the title compound as ayellow gum (320 mg).

¹H-NMR (400 MHz, DMSO-d₆) δ ppm: 7.67 (2H, m), 7.25 (1H, t), 7.00 (2H,m), 6.68 (1H, dd), 6.53 (1H, t), 6.49 (1H, dd), 3.73 (3H, s), 3.39-3.46(1H, m), 1.22 (3H, d); UPLC: 0.51 min, 287 [M+H]+.

Intermediate 3 1-(ethyloxy)-3-[(4-nitrophenyl)oxy]benzene

The two reactions were performed in parallel. Two microwave vials wereset up in parallel. In a large 30 mL microwave vial, 3-(ethyloxy)phenol(2 times 1.25 g, 9.045 mmol) was dissolved in 6 mL of dimethylformamide.1-Fluoro-4-nitrobenzene (2 times 1.28 g, 9.045 mmol) and potassiumcarbonate (2 times 3.75 g, 27.15 mmol) were added. The reaction mixturewas heated under microwave irradiation during 1 hour at 120° C. Thecombined reaction mixtures were filtered. The filtrated solid was washedwith dichloromethane. The volatiles were evaporated under vacuum. Somedichloromethane and brine were added to this crude. The compound wasextracted with dichloromethane (2 times) and ethyl acetate (2 times).The combined organic phases were dried over sodium sulphate. Thisafforded the title compound (4 g).

¹H NMR (400 MHz, methanol-d₄): 6 ppm 8.38-8.05 (2H, m), 7.46-7.31 (1H,m), 7.20-7.05 (2H, m), 6.94-6.81 (1H, m), 6.77-6.63 (2H, m), 4.08 (2H,q), 1.42 (3H, t); UPLC: 0.89 min, 260 [M+H]+.

Intermediate 4 4-{[3-(ethyloxy)phenyl]oxy}aniline

A solution of 1-(ethyloxy)-3-[(4-nitrophenyl)oxy]benzene (Intermediate3, 4 g) and tin chloride monohydrate (28.8 g, 139 mmol) in ethyl acetate(200 mL) was heated at reflux overnight (15 hours). The reaction mixturewas cooled down. It was then diluted with ethyl acetate (50 mL), washedwith saturated NaHCO3 (100 mL), brine (100 mL) and dried over sodiumsulphate. After evaporation of the volatiles, the residue was purifiedby an SCX (wash of the column with methanol, adsorption of the compound,wash with methanol (3CV), desorption with 2N methanolic ammonia (3CV)).Evaporation afforded the title compound (2.9 g).

¹HNMR (400 MHz, CDCl₃): δ ppm 6.50 (1H, t), 6.21 (2H, d), 6.06 (2H, d),5.91-5.84 (3H, m), 3.34-3.29 (4H, m), 0.72 (3H, t); UPLC: 0.89 min, 260[M+H]+.

Intermediate 5 3-chloro-5-fluorophenyl 4-nitrophenyl ether

To a solution of 3-chloro-5-fluorophenol (1.46 g, 10 mmol) and1-fluoro-4-nitrobenzene (1.41 g, 10 mmol) in acetonitrile (40 mL) wasadded potassium carbonate (2.76 g, 20 mmol) and the reaction mixture washeated to reflux for 4 hours. After filtration, the solvent was removed.The residue obtained was washed with n-hexane (2 times 15 mL) and driedto afford the title compound (2.38 g) which was directly used in thenext step.

Intermediate 6 4-[(3-chloro-5-fluorophenyl)oxy]aniline

To a solution of 3-chloro-5-fluorophenyl 4-nitrophenyl ether(Intermediate 5, 2.38 g) in THF (40 mL) and water (10 mL) was added Fepower (11.2 g, 200 mmol) and ammonium chloride (10.7 g, 200 mmol). Thereaction mixture was heated at reflux for 4 hours. After filtration, thesolvent was concentrated to give a residue and poured into 50 mL ofwater. The mixture was extracted with ethyl acetate (3 times 50 mL) andthe combined organic phases were washed and dried over magnesiumsulphate. Removal of the solvent afforded the title compound (2.02 g)which was directly used in the next step.

Intermediate 7 3-chloro-4-fluorophenyl 4-nitrophenyl ether

To a solution of 3-chloro-4-fluorophenol (1.46 g, 10 mmol) and1-fluoro-4-nitrobenzene (1.41 g, 10 mmol) in acetonitrile (40 mL) wasadded potassium carbonate (2.76 g, 20 mmol). The reaction mixture washeated at reflux for 4 hours. After filtration, the solvent was removedto give a residue. The residue was washed with n-hexane (2 times 15 mL)and dried to afford the title compound (2.48 g) which was directly usedin the next step.

Intermediate 8 4-[(3-chloro-4-fluorophenyl)oxy]aniline

To a solution of 3-chloro-4-fluorophenyl 4-nitrophenyl ether(Intermediate 7, 2.48 g) in THF/water (40 mL/10 mL) was added Fe power(11.2 g, 200 mmol) and ammonium chloride (10.7 g, 200 mmol and themixture was heated at reflux for 4 hours. After filtration, the solventwas concentrated to give a residue and poured into 50 mL of water. Themixture was extracted with ethyl acetate (3 times 50 mL) and thecombined organic phases were washed and dried over magnesium sulphate.Removal of the solvent afforded the title compound (2.15 g) which wasdirectly used in the next step.

Intermediate 9N-[({4-[(3-chloro-4-fluorophenyl)oxy]phenyl}amino)carbonyl]-D-alanine

To a solution of 4-[(3-chloro-4-fluorophenyl)oxy]aniline (Intermediate8, 237 mg) and triphosgene (99 mg, 0.33 mmol) in 15 mL ofdichloromethane was added DIPEA (155 mg, 1.2 mmol) and the mixture wasstirred at room temperature for 2 hours. Then the solvent was evaporatedto give a residue. The residue was dissolved in 5 mL of THF and wastransferred to a mixture of DIPEA (65 mg, 0.5 mmol, Acros) and D-Alanine(89 mg, 1 mmol) in 5 mL of THF. The whole reaction mixture was stirredat room temperature for 16 hours. Removal of the solvent afforded thetitle compound (352 mg) which was directly used in the next step.

MS_(—)2 (ESI): 353 [M+H]+

Intermediate 10N-[({4-[(3-chloro-4-fluorophenyl)oxy]phenyl}amino)carbonyl]-L-alanine

The title compound was made in a similar fashion to the preparation ofintermediate 9 replacing D-alanine with L-alanine (89 mg, 1 mmol) toafford the title compound (325 mg), which was used directly in the nextstep.

MS_(—)2 (ESI): 353 [M+H]+

Intermediate 11 2-methyl-5-(methyloxy)aniline

A suspension of 1-methyl-4-(methyloxy)-2-nitrobenzene (20.0 g, 119.8mmol) and Pd/C (10%, 3 g) in methanol (100 mL) was stirred under H2atmosphere at room temperature overnight. The mixture was filteredthrough a pad of celite and the filtrate was evaporated under vacuum toafford the title compound as a solid (16.1 g).

MS_(—)2 (ESI): 138 [M+H]+.

Intermediate 12 2-methyl-5-(methyloxy)phenol

To a solution of 2-methyl-5-(methyloxy)aniline (Intermediate 11, 6.0 g)in H2SO4 (5 M, 20 mL) was added portionwise NaNO2 (3.4 g, 49.3 mmol) at0-5° C. The mixture was stirred at 50° C. for 1 hour and extracted withethyl acetate (4 times 30 mL). The combined ethyl acetate layers weredried over sodium sulphate and concentrated under vacuum to give aresidue, which was purified by column chromatography on silica gel (withEtOAc:PE=1:20 as eluents) to afford the title compound as a solid.

MS_(—)2 (ESI): 139 [M+H]+.

Intermediate 13 1-methyl-4-(methyloxy)-2[(4-nitrophenyl)oxy]benzene

To a solution of 2-methyl-5-(methyloxy)phenol (Intermediate 12, 1.5 g)and 1-fluoro-4-nitrobenzene (1.4 g, 10.0 mmol) in acetonitrile (100 mL)was added potassium carbonate (2.1 g, 15.2 mmol) and the mixture wasstirred at reflux for 5 hours. The resulting mixture was concentratedand partitioned between ethyl acetate (3 times 30 mL) and water (100mL). The combined ethyl acetate layers were dried over sodium sulphate,filtered and concentrated under vacuum. The crude product thus obtainedwas purified by column chromatography on silica gel (EtOAc: PE=1:20) toafford the title compound as a solid (2.5 g).

Intermediate 14 4-{[2-methyl-5-(methyloxy)phenyl]oxy}aniline

A suspension of 1-methyl-4-(methyloxy)-2-[(4-nitrophenyl)oxy]benzene(Intermediate 13, 2.5 g) and Pd/C (10%, 1 g) in MeOH (100 mL) wasstirred under H2 atmosphere for overnight at room temperature andfiltered through a pad of celite. The filtrate was evaporated to affordthe title compound as a solid (2.0 g).

¹HNMR (400 MHz, CDCl₃): δ ppm 7.14-7.12 (1H, d), 6.85-6.82 (2H, d),6.68-6.66 (2H, d), 6.59-6.56 (1H, d), 6.40 (1H, s), 3.74-3.71 (5H, m),2.25 (3H, s); MS_(—)2 (ESI): 230 [M+H]+

Intermediate 15 1,1-dimethylethyl{(1R)-1-methyl-2-[(4-{[2-methyl-5-(methyloxy)phenyl]oxy}phenyl)amino]-2-oxoethyl}carbamate

To a solution of N-{[(1,1-dimethylethyl)oxy]carbonyl}-D-alanine (89 mg,0.471 mmol) in dry N,N-dimethylformamide (5 mL), DIPEA (0.103 mL, 0.589mmol) then HATU (179 mg, 0.471 mmol) were added and the reaction mixturewas stirred for 15 minutes at room temperature under argon. Then4-{[2-methyl-5-(methyloxy)phenyl]oxy}aniline (Intermediate 14, 90 mg)was added and the reaction mixture was stirred at 60° C. under argon for1 hour 30 min. The reaction mixture was evaporated. The residuesobtained was purified by silica gel chromatography (Companioninstrument, 40 g silica cartridge) eluting with a gradient cHex/EtOAc100/0 to 75/25 during 15 min and then 75/25 during 30 min to afford thetitle compound (155 mg).

¹H-NMR (400 MHz, methanol-d₄): 6 ppm 7.52 (2H, d), 7.13 (1H, d), 6.85(2H, d), 6.64 (1H, dd), 6.42 (1H, d), 4.22 (1H, dd), 3.57-3.78 (3H, m),2.12 (3H, s), 1.46 (9H, s), 1.41 (3H, d);

UPLC_B: 1.04 min, 401 [M+H]+.

Intermediate 16N¹-(4-{[2-methyl-5-(methyloxy)phenyl]oxy}phenyl)-D-alaninamide

1,1-dimethylethyl{(1R)-1-methyl-2-[(4-{[2-methyl-5-(methyloxy)phenyl]oxy}phenyl)amino]-2-oxoethyl}carbamate(Intermediate 15, 150 mg) was dissolved in 3 mL of dry dichloromethane.To this solution at 0° C. under argon was added dropwise 30 equivalentsof TFA (0.866 mL, 11.24 mmol). The reaction was stirred at 0° C. for 4hours. The reaction mixture was evaporated. The residue obtained waspurified with an SCX cartridge (the cartridge was washed with 3 CV ofmethanol, then the compound was adsorbed on the cartridge, washed with 5CV of methanol and desorbed with 2 CV of methanolic ammonia (1N)).Evaporation of the volatiles, afforded the title compound (129 mg).

¹H-NMR (400 MHz, methanol-d₄): δ ppm 7.48 (2H, m), 7.10 (1H, d), 6.81(2H, m), 6.60 (1H, dd), 6.36 (1H, d), 3.64 (3H, s), 3.54 (1H, m), 2.06(3H, s), 1.33 (3H, d); UPLC_B: 0.77 min, 301 [M+H]+.

Intermediate 17 4-methyl-3-(methyloxy)aniline

To a solution of 1-methyl-2-(methyloxy)-4-nitrobenzene (2.5 g, 14.96mmol) in methanol (50 mL) Ni-Raney (˜2 g) was added and the reactionmixture was stirred overnight at room temperature under H2 atmosphere (1atm). The catalyst was filtered off and the residue was purified by SCXcartridge (50 g) to afford the title compound (1.86 g) as a colourlessoil.

¹H-NMR (400 MHz, DMSO-d₆) δ ppm: 6.73 (1H, d), 6.19 (1H, d), 6.05 (1H,dd), 4.85 (2H, s), 3.68 (3H, s), 1.97 (3H, s); UPLC_B: 0.62 min, 138[M+H]+.

Intermediate 18 4-methyl-3-(methyloxy)phenol

To a suspension of 4-methyl-3-(methyloxy)aniline (Intermediate 17, 1.86g) in water (100 mL)/H2SO4 (30 mL, 563 mmol) at 0° C. a solution ofsodium nitrite (1.029 g, 14.91 mmol) in water (10 mL) was slowly addedand the reaction mixture was stirred for 30 minutes at 0° C. Thereaction mixture was slowly added to a solution of H2SO4 98% (20 mL) inWater (80 mL) pre-heated at 90° C. and stirred at this temperature for 1h. After cooling the mixture was extracted with Et2O (2×200 mL), theorganic layer was dried on sodium sulphate, filtered and evaporated toafford the title compound (1.86 g) as a red/brown oil.

¹H-NMR (400 MHz, DMSO-d₆) δ ppm: 9.14 (1H, br.s), 6.87 (1H, d), 6.35(1H, d), 6.24 (1H, dd), 3.71 (3H, s), 2.01 (3H, s); UPLC_B: 0.63 min,137 [M−H]−.

Intermediate 19 1-methyl-2-(methyloxy)-4-[(4-nitrophenyl)oxy]benzene

To a solution of 4-methyl-3-(methyloxy)phenol (Intermediate 18, 0.800 g)in dry acetonitrile (60 mL) potassium carbonate (1.600 g, 11.58 mmol)and then 1-fluoro-4-nitrobenzene (817 mg, 5.79 mmol) were added and thereaction mixture was refluxed for 6 hours. The solid was filtered offand the solvent evaporated affording the title compound (1.43 g) as anorange solid.

¹H-NMR (400 MHz, DMSO-d₆) δ ppm: 8.24 (2H, m), 7.23 (1H, d), 7.11 (2H,m), 6.82 (1H, d), 6.66 (1H, dd), 3.78 (3H, s), 2.16 (3H, s); UPLC_B:1.03 min, 260 [M+H]+.

Intermediate 20 4-{[4-methyl-3-(methyloxy)phenyl]oxy}aniline

To a solution of 1-methyl-2-(methyloxy)-4-[(4-nitrophenyl)oxy]benzene(Intermediate 19, 1.43 g) in tetrahydrofuran (65 mL)/water (32.5 mL)iron (1.540 g, 27.6 mmol) and then ammonium chloride (1.475 g, 27.6mmol) were added and the reaction mixture was stirred for 5 hours atroom temperature. The catalyst was filtered off and the solution wasdiluted with a saturated solution of Na2CO3 (10 mL) and extracted withethyl acetate (2 times 60 mL). Combined organic layers were dried oversodium sulphate, filtered and evaporated to the title compound (1.25 g)as a brown/red solid.

¹H-NMR (400 MHz, DMSO-d₆) δ ppm: 7.00 (1H, d), 6.77-6.70 (2H, m),6.60-6.51 (3H, m), 6.24 (1H, dd), 4.94 (2H, br. s), 3.71 (3H, s), 2.06(3H, s); UPLC_B: 0.86 min, 230 [M+H]+.

Intermediate 21 1,1-dimethylethyl{(1R)-1-methyl-2-[(4-{[4-methyl-3-(methyloxy)phenyl]oxy}phenyl)amino]-2-oxoethyl}carbamate

To a solution of N-{[(1,1-dimethylethyl)oxy]carbonyl}-D-alanine (182 mg,0.960 mmol) in dry N,N-Dimethylformamide (DMF) (4 mL) DIPEA (0.305 mL,1.745 mmol) and then TBTU (336 mg, 1.047 mmol) were added and thereaction mixture was stirred for 15 minutes at r.t.4-{[4-methyl-3-(methyloxy)phenyl]oxy}aniline (Intermediate 20, 200 mg)was then added and the reaction mixture was stirred for 1 hour at thesame temperature. The reaction was quenched with water (2 mL), dilutedwith brine (10 mL) and extracted with ethyl acetate (2 times 20 mL).Organic layer was dried over sodium sulphate, filtered and evaporatedand the residue was purified by flash chromatography (Biotage system, 10g SNAP column) using as eluent a gradient cyclohexane/ethyl acetate from100/0 to 80/20 to afford the title compound as a yellow pale solid (304mg).

¹H-NMR (400 MHz, DMSO-d₆) δ ppm: 9.93 (s, 1H), 7.60 (m, 2H), 7.09 (d,2H), 6.97 (m, 2H), 6.63 (d, 1H), 6.39 (dd, 1H), 4.15-4.03 (m, 1H), 3.74(s, 3H), 2.10 (s, 3H), 1.39 (s, 9H), 1.26 (d, 3H), UPLC_B: RT 0.96 min,m/z 401 [M+H]+.

Intermediate 22N¹-(4-{[4-methyl-3-(methyloxy)phenyl]oxy}phenyl)-D-alaninamide

To a solution of 1,1-dimethylethyl{(1R)-1-methyl-2-[(4-{[4-methyl-3-(methyloxy)phenyl]oxy}phenyl)amino]-2-oxoethyl}carbamate(Intermediate 21, 300 mg) in dry dichloromethane (7.5 mL) TFA (2.5 mL,32.4 mmol) was slowly added and the reaction mixture was stirred for 1.5hours at room temperature. The solvent and the excess of TFA wereevaporated and the residue was purified by SCX cartridge (10 g) toafford the title compound as an orange oil (219 mg).

¹H-NMR (400 MHz, DMSO-d₆) δ ppm: 7.64 (2H, m), 7.08 (1H, s), 6.96 (2H,m), 6.63 (1H, d), 6.39 (1H, dd), 3.74 (3H, s), 3.41 (1H, q), 2.10 (3H,s), 1.21 (3H, d); UPLC_B: 0.80 min, 301 [M+H]+.

Intermediate 23 2-{[3-(1-methylethyl)phenyl]oxy}-5-nitropyridine

In a 30 mL large microwave vial, 2-chloro-5-nitropyridine (1.041 g, 6.57mmol, 1 equiv) was dissolved in 5.5 mL of dimethylformamide.3-(1-methylethyl)phenol (0.90 mL, 6.57 mmol, 1 equiv) and potassiumcarbonate (4.54 g, 32.8 mmol, 5 equiv) were added. The reaction mixturewas heated under microwave irradiation for 1 hour at 110° C. (BiotageInitiator). The reaction mixture was filtered. The filtrated solid waswashed with dichloromethane (30 mL). The volatiles were evaporated undervacuum. The crude compound was dissolved in dichoromethane (20 mL) andbrine was added (20 mL). The compound was extracted 2 times withdichloromethane (2×20 mL) and 2 times with ethyl acetate (2×20 mL). Theorganic phase was dried over sodium sulphate. Evaporation afforded thetitle compound (1.402 g).

¹HNMR (400 MHz, methanol-d₄): δ ppm 8.94 (1H, d), 8.52 (1H, dd), 7.33(1H, t), 7.15 (1H, d), 7.06 (1H, d), 7.02 (1H, t), 6.90-6.97 (1H, m),2.81 (3H, s); UPLC: 0.93 min, 259 [M+H]+.

Intermediate 24 6-{[3-(1-methylethyl)phenyl]oxy}-3-pyridinamine

2-{[3-(1-methylethyl)phenyl]oxy}-5-nitropyridine (Intermediate 23, 1.39g) was dissolved in ethanol (25 mL). Hydrazine monohydrate (0.524 mL,1076 mmol) and palladium on carbon (401 mg, 0.377 mmol) were added. Thereaction mixture was heated at reflux under argon for 1 hour. Thereaction was cooled down and then filtered on celite. The organic phasewas evaporated under vacuum. The residue was purified by flashchromatography on silica (Companion instrument, 120 g silica cartridge,gradient cyclohexane/ethylacetate from 100/0 to 30/70 in 15 min then30/70 during 30 min). Evaporation afforded the title compound as ayellow oil (821 mg).

¹HNMR (400 MHz, methanol-d₄): δ ppm 7.65 (1H, d), 7.29-7.15 (2H, m),6.99 (1H, d), 6.81-6.86 (1H, m), 6.68-6.78 (2H, m), 2.97-2.75 (1H, m),1.23 (3H, s), 1.22 (3H, s); UPLC: 0.70 min, 229 [M+H]+.

Intermediate 251,1-dimethylethyl{(1R)-1-methyl-2-[(6-{[3-(1-methylethyl)phenyl]oxy}-3-pyridinyl)amino]-2-oxoethyl}carbamate

To a solution of N-{[(1,1-dimethylethyl)oxy]carbonyl}-D-alanine (69.6mg, 0.368 mmol) in dry N,N-dimethylformamide (4 mL), DIPEA (0.080 mL,0.460 mmol), HATU (140 mg, 0.368 mmol) were added and the reactionmixture was stirred for 15 minutes at room temperature under argon. Then6-{[3-(1-methylethyl)phenyl]oxy}-3-pyridinamine (Intermediate 24, 70 mg)was added and the reaction mixture was stirred overnight at 60° C. underargon. The reaction mixture was evaporated. The residue obtained waspurified by silica gel chromatography (Companion system, 12 g cartridge)with a gradient cyclohexane/ethyl acetate from 100/0 to 65/35 to affordthe title compound (59 mg).

¹HNMR (400 MHz, methanol-d₄): δ ppm 8.33 (1H, d), 8.05 (1H, dd), 7.30(1H, t), 7.08 (1H, d), 6.92-6.98 (1H, m), 6.83-6.91 (2H, m), 4.12-4.29(1H, m), 2.79-2.97 (1H, m), 1.45 (9H, s), 1.38 (3H, d), 1.25 (6H, d);UPLC: 0.85 min, 400 [M+1]+

Intermediate 26N¹-(6-{[3-(1-methylethyl)phenyl]oxy}-3-pyridinyl)-D-alaninamide

1,1-dimethylethyl{(1R)-1-methyl-2-[(6-{[3-(1-methylethyl)phenyl]oxy}-3-pyridinyl)amino]-2-oxoethyl}carbamate(Intermediate 25, 56 mg) was dissolved in 3 mL of dry dichloromethane.To this solution at 0° C. were added dropwise 30 equivalents of TFA(0.324 mL). The reaction was stirred at 0° C. for 3 hours. The reactionmixture was evaporated. The crude obtained was purified by SCX on a 5 gcartridge. 3 CV of methanol were used first, then the residue wasadsorbed on the cartridge, washed with 5 CV of methanol and desorbedwith 2 CV of methanolic ammonia (1N). Evaporation of the volatiles,afforded the title compound (38 mg).

¹HNMR (400 MHz, methanol-d₄): δ ppm 8.40 (1H, d), 8.10 (1H, dd), 7.33(1H, t), 7.10 (1H, d), 6.98 (1H, t), 6.85-6.92 (2H, m), 3.52-3.69 (1H,m), 2.78-3.02 (1H, m), 1.39 (3H, d), 1.27 (6H, d); UPLC: 0.59 min, 300[M+1]+

Intermediate 27 3-[(1-methylethyl)oxy]phenol

To a solution of 1,3-benzenediol (8 g, 72.7 mmol) and 2-iodopropane (12g, 70.6 mmol) in ethanol (100 mL preheated at reflux) was added asolution of KOH (83%, 5.3 g, 77.6 mmol) in water (20 mL) over a periodof 30 minutes. The mixture was refluxed for 3 hours and poured into NaOH(1N, 100 mL). The resulting mixture was extracted with ethyl acetate (3times 50 mL) and the aqueous layer was acidified with 10% HCl to adjustthe pH=5 and extracted with ethyl acetate (3 times 50 mL). The combinedextracts were washed with brine (50 mL), dried and concentrated undervacuum. The residue was purified by column chromatography on silica gel(PE: EtOAc=5:1) to afford the title compound as a colourless oil (2.1g).

MS 1 (ESI): 151 [M−H]−

Intermediate 28 2-({3-[(1-methylethyl)oxy]phenyl}oxy)-5-nitropyridine

To a solution of 3-[(1-methylethyl)oxy]phenol (Intermediate 27, 456 mg)in DMSO (8 mL) was added t-BuOK (336 mg, 3 mmol, Acros). The reactionmixture was stirred at 20° C. for 30 minutes. 2-chloro-5-nitropyridine(474 mg, 3 mmol, Aldrich) was added and the resulting mixture wasstirred at 120° C. for 2 hours. The reaction mixture was cooled to roomtemperature, poured into ice-water (50 mL) and extracted withdichloromethane (3 times 50 mL). The combined organic layers were driedover sodium sulphate, filtered and concentrated. The residue waspurified by column chromatography on silica gel elutiong with (PE:EtOAc=50:1) to afford the title compound as a light yellow solid (670mg).

MS 1 (ESI): 275 [M+H]+

Intermediate 29 6-({3-[(1-methylethyl)oxy]phenyl}oxy)-3-pyridinamine

To a solution of 2-({3-[(1-methylethyl)oxy]phenyl}oxy)-5-nitropyridine(Intermediate 28, 670 mg, 2.45 mmol) in methanol (50 mL) was added Pd/C(10%, 100 mg, 0.1 wet. e.g.) and the flask was filled in with H₂. Theresulting mixture was stirred at room temperature under H₂ atmosphereovernight and filtered. The filtrate was concentrated under vacuum toafford the title compound as a brown solid (560 mg).

MS 1 (ESI): 245 [M+H]+

Intermediate 30 N-{[(1,1-dimethylethyl)oxy]carbonyl}-D-alanine

To a solution of D-alanine (4.45 g, 50 mmol) in THF (100 mL) and water(50 mL) was added a solution of NaHCO3 (4.2 g, 50 mmol) in water (30mL). After stirring for 15 minutes, a solution of Boc-anhydride (16.35g, 75 mmol) in THF (20 mL) was added and the mixture was stirred at roomtemperature for 4 hours. The solvent was evaporated and 2N HCl was usedto adjust the pH=3-4. The mixture was extracted with ethyl acetate (3times 200 mL) and the combined ethyl acetate layers were washed withbrine (50 mL), dried and concentrated. The residue was recrystallizedwith ethyl acetate/hexane to afford the title compound as a white solid(5 g).

¹HNMR (DMSO-d₆): δ ppm 12.38 (1H, s), 7.11-7.09 (1H, d), 3.94-3.88 (1H,m), 1.38 (9H, s), 1.22-1.21 (3H, d).

Intermediate 311,1-dimethylethyl((1R)-1-methyl-2-{[6-({3-[(1-methylethyl)oxy]phenyl}oxy)-3-pyridinyl]amino}-2-oxoethyl)carbamate

A solution of 6-({3-[(1-methylethyl)oxy]phenyl}oxy)-3-pyridinamine(Intermediate 29, 244 mg, 1 mmol),N-{[(1,1-dimethylethyl)oxy]carbonyl}-D-alanine (Intermediate 30, 284mg), HBTU (567 mg, 1.5 mmol) and DIPEA (194 mg, 1.5 mmol, Acros) in DMF(8 mL) was heated under microwave (Biotage instrument) at 110° C. for 3hours. The solvent was distilled off to afford the title compound as abrown oil, which was used directly in the next step (400 mg, 96% yield).

MS_(—)2 (ESI): 416 [M+H]+

Intermediate 32N¹-[6-({3-[(1-methylethyl)oxy]phenyl}oxy)-3-pyridinyl]-D-alaninamide

To a solution of 1,1-dimethylethyl((1R)-1-methyl-2-{[6-({3-[(1-methylethyl)oxy]phenyl}oxy)-3-pyridinyl]amino}-2-oxoethyl)carbamate(Intermediate 31, 400 mg, 0.96 mmol) in dichloromethane (14 mL) wasadded TFA (6 mL) portionwise during 15 minutes at 0° C. The resultingmixture was stirred at room temperature for 2 hours. The solvent wasdistilled off to afford the title compound (260 mg, 85%) as a grey oil.

MS 1 (ESI): 316 [M+H]+

Intermediate 33 2-[(2,5-dimethylphenyl)oxy]-5-nitropyridine

In a microwave vial, 2-chloro-5-nitropyridine (80 mg, 0.505 mmol) wasdissolved in 2 mL of dry dimethylformamide. 2,5-dimethylphenol (80 mg,0.505 mmol, 1 equiv) and potassium carbonate (418 mg, 3.03 mmol, 6equiv) were added. The reaction mixture was heated under microwaveirradiation for 1 hour at 110° C. (Biotage Initiator). The reactionmixture was filtered. The filtrated solid was washed withdichloromethane (5 mL). The volatiles were evaporated. The residue wasdissolved in dichoromethane (10 mL) and brine was added (10 mL). Theorganic layer was extracted 2 times with dichloromethane (2×15 mL) and 2times with ethylacetate (2×15 mL). The organic phase was dried oversodium sulphate. The solvent was removed to afford the title compound(112 mg)

¹HNMR (400 MHz, methanol-d₄): δ ppm 8.97 (1H, d), 8.58 (1H, dd),7.20-6.90 (4H, m), 2.32 (3H, s), 2.07 (3H, s); UPLC: 0.87 mins, 245[M+H]+.

Intermediate 34 6-[(2,5-dimethylphenyl)oxy]-3-pyridinamine

2-[(2,5-dimethylphenyl)oxy]-5-nitropyridine (Intermediate 33, 140 mg,0.450 mmol) was dissolved in ethanol (3 mL). Hydrazine hydrate (83 μL,0.884 mmol) and palladium on carbon (47 mg, 0.044 mmol) were added. Thereaction mixture was heated at reflux under argon. After overnightheating, the reaction was cooled down. The reaction mixture wasfiltered. The organic phase was evaporated under vacuum. The residue waspurified by SCX (wash with MeOH, desorbed with 2N methanolic ammonia).Evaporation afforded the title compound (85 mg).

UPLC: 0.68 min, 215 [M+H]+.

Intermediate 35 1,1-dimethylethyl[(1R)-2-({6-[(2,5-dimethylphenyl)oxy]-3-pyridinyl}amino)-1-methyl-2-oxoethyl]carbamate

The title compound was made in a similar fashion to the preparation ofIntermediate 25 replacing6-{[3-(1-methylethyl)phenyl]oxy}-3-pyridinamine with6-[(2,5-dimethylphenyl)oxy]-3-pyridinamine (Intermediate 34) and usingthe following conditions for the silica gel chromatography: Companioninstrument, 12 g cartridge, a gradient cHex/EtOAc as eluent from 100/0to 70/30. This afforded the title compound as a light brown oil (63 mg).

¹HNMR (400 MHz, methanol-d₄): δ ppm 8.31 (1H, d), 8.05 (1H, dd), 7.18(1H, d), 6.98 (1H, d), 6.76-6.87 (2H, m), 4.02-4.34 (1H, m), 2.35 (3H,s), 2.12 (3H, s) 1.47 (9H, s) 1.41 (3H, d);

UPLC: 0.84 min, 386 [M+1]+

Intermediate 36N¹-{6-[(2,5-dimethylphenyl)oxy]-3-pyridinyl}-D-alaninamide

1,1-dimethylethyl[(1R)-2-({6-[(2,5-dimethylphenyl)oxy]-3-pyridinyl}amino)-1-methyl-2-oxoethyl]carbamate(Intermediate 35, 60 mg) was dissolved in 4 mL of dry dichloromethane.To this solution, at 0° C., were added dropwise 40 equivalents of TFA(0.480 mL). The reaction was stirred for 3 hours 30 at 0° C. Thereaction mixture was evaporated and then purified by SCX on a 5 gcartridge. 3 CV of methanol were used first, then the residue wasadsorbed on the cartridge, washed with 5 CV of methanol and desorbedwith CV of methanolic ammonia (1N). Evaporation of the volatiles,afforded the title compound (49 mg)

¹HNMR (400 MHz, methanol-d₄): δ ppm 8.34 (1H, d), 8.07 (1H, dd), 7.18(1H, d), 6.99 (1H, d), 6.69-6.90 (2H, m), 3.60 (1H, q), 2.32 (3H, s),2.12 (3H, s), 1.39 (3H, d); UPLC: 0.54 min, 286 [M+1]+

Intermediate 37 2-[(2,3-dimethylphenyl)oxy]-5-nitropyridine

In a 20 mL microwave vial 2-chloro-5-nitropyridine (500 mg, 3.15 mmol),2,3-dimethylphenol (385 mg, 3.15 mmol) and potassium carbonate (1308 mg,9.46 mmol) were dissolved in N,N-Dimethylformamide (10 mL) to give adark brown suspension. The reaction vessel was sealed and heated inBiotage Initiator at 110° C. for 1 h. After cooling the reaction wasdiluted with 25 mL of Et₂O. The organic phase was washed with 3×25 mL ofwater, 10 mL of brine, dried over sodium sulphate, filtered andevaporated under vacuum to afford the title compound as a pale orangeoil. (640 mg). ¹H NMR (400 MHz, CDCl₃): δ ppm 9.07 (1H, d), 8.50 (1H,dd), 7.24-7.19 (1H, m), 7.18-7.14 (1H, m), 7.04 (1H, d), 6.95 (1H, d),2.38 (3H, s), 2.09 (3H, s); UPLC: 0.81 min, 245 [M+H]+.

Intermediate 38 6-[(2,3-dimethylphenyl)oxy]-3-pyridinamine

In a 50 mL round-bottomed flask2-[(2,3-dimethylphenyl)oxy]-5-nitropyridine (Intermediate 37, 640 mg)was dissolved in ethanol (10 mL) to give a pale yellow solution.Hydrazine hydrate (0.463 mL, 4.72 mmol) and palladium on carbon (25.10mg, 0.236 mmol) were added. The reaction mixture was stirred at 90° C.After 1 hour, the reaction was complete. The reaction mixture wasfiltered and the organic phase was evaporated in vacuo affording thetitle compound as a pale yellow oil (573 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 7.72 (1H, d), 7.05-7.16 (2H, m), 7.01(1H, d), 6.86 (1H, d), 6.71 (1H, d), 3.48 (2H, br. s), 2.34 (3H, s),2.16 (3H, s); UPLC: 0.62 min, 215 [M+H]+.

Intermediate 391,1-dimethylethyl[(1R)-2-({6-[(2,3-dimethylphenyl)oxy]-3-pyridinyl}amino)-1-methyl-2-oxoethyl]carbamate

To a solution of N-{[(1,1-dimethylethyl)oxy]carbonyl}-D-alanine (26.5mg, 0.140 mmol) in dry N,N-dimethylformamide (4 mL), DIPEA (31 μl, 0.175mmol, 1.5 equiv) and then HATU (53.2 mg, 0.140 mmol, 1.2 equiv) wereadded and the reaction mixture was stirred for 15 minutes at roomtemperature under argon. Then 6-[(2,3-dimethylphenyl)oxy]-3-pyridinamine(Intermediate 38, 25 mg) was added and the reaction mixture was stirredat 60° C. under argon. The reaction mixture was left overnight underheating. It was then was evaporated. The residue obtained was directlypurified on silica gel chromatography (Companion instrument, 2×4 gcartridge) with a gradient cyclohexane/ethylacetate 100/0 to 70/30during 15 min and 70/30 during 20 min. This afforded the title compound(31 mg).

¹HNMR (400 MHz, methanol-d₄): δ ppm 8.29 (1H, d), 8.02 (1H, dd),7.20-6.99 (3H, m), 6.81 (2H, dd), 4.21 (1H, m), 2.32 (3H, s), 2.08 (3H,s), 1.45 (9H, s), 1.40 (3H, d); UPLC: 0.80 min, 386 [M+1]+

Intermediate 40N¹-{6-[(2,3-dimethylphenyl)oxy]-3-pyridinyl}-D-alaninamide

1,1-dimethylethyl[(1R)-2-({6-[(2,3-dimethylphenyl)oxy]-3-pyridinyl}amino)-1-methyl-2-oxoethyl]carbamate(Intermediate 39, 29 mg) was dissolved in 3 mL of dry dichloromethane.To this solution at 0° C. under argon was added dropwise 30 equivalentsof TFA (168 μl, 2.179 mmol). The reaction was stirred during 1 hour at0° C. and 2 hours at room temperature. The reaction mixture wasevaporated. The residue obtained was purified by SCX (3 CV of methanolwere used first, then the residue was adsorbed on the cartridge, washedwith 5 CV of methanol and desorbed with 2 CV of methanolic ammonia(1N)). Evaporation of the volatiles, afforded the title compound (21mg).

UPLC: 0.52 min, 286 [M+1]+

Intermediate 41 2-[(2,6-dimethylphenyl)oxy]-5-nitropyridine

In a 20 mL microwave vial 2-chloro-5-nitropyridine (500 mg, 3.15 mmol)was dissolved in N,N-dimethylformamide (10 mL) to give a pale yellowsolution. 2,6-dimethylphenol (385 mg, 3.15 mmol) and potassium carbonate(1308 mg, 9.46 mmol) were added. The reaction vessel was sealed andheated under microwave irradiation (Biotage instrument) at 110° C. for 1hour. The reaction mixture was quenched with 10 mL of water and dilutedwith 10 mL of Et₂O. Phases were separated through a separating funnel.The organic phase was washed with 3 times 10 mL of water, 10 mL ofbrine, dried over sodium sulphate, filtered and evaporated under vacuumto afford the title compound as a pale orange oil (555.9 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 9.04 (1H, d), 8.50 (1H, dd), 7.16 (3H,s), 7.03 (1H, d), 2.12 (6H, s); UPLC_B: 0.95 mins, 245 [M+H]+.

Intermediate 42 6-[(2,6-dimethylphenyl)oxy]-3-pyridinamine

In a 50 mL round-bottomed flask2-[(2,6-dimethylphenyl)oxy]-5-nitropyridine (Intermediate 41, 555.9 mg)was dissolved in ethanol (10 mL) to give a pale orange solution.Palladium on carbon (230 mg, 0.216 mmol) and hydrazine hydrate (0.416mL, 4.32 mmol) were added. The reaction mixture was stirred at 90° C.After 3 hours the reaction was completed. The reaction mixture wasfiltered and the organic phase was evaporated under vacuum to afford929.9 mg of a dark orange solid that was charged on a 10 g SCXcartridge. It was then flushed with 200 mL of ethanol followed by 50 mLof 2M solution of ammonia in MeOH. The ammonia eluate was evaporatedunder vacuum to afford the title compound as dark orange solid (447.6mg).

¹H NMR (400 MHz, CDCl₃): d ppm 7.64 (1H, d), 7.02-7.12 (4H, m), 6.62(1H, d), 3.50 (2H, br. s), 2.14 (6H, s); UPLC_B: 0.74 mins, 215 [M+H]+.

Intermediate 431,1-dimethylethyl[(1R)-2-({6-[(2,6-dimethylphenyl)oxy]-3-pyridinyl}amino)-1-methyl-2-oxoethyl]carbamate

In a 8 mL vial N-{[(1,1-dimethylethyl)oxy]carbonyl}-D-alanine (190 mg,1.003 mmol) was dissolved in N,N-dimethylformamide (4 mL) to give acolourless solution. N-ethyl-N-(1-methylethyl)-2-propanamine (0.219 mL,1.253 mmol) andN-[(dimethylamino)(3H-[1,2,3]triazolo[4,5-b]pyridin-3-yloxy)methylidene]-N-methylmethanaminiumhexafluorophosphate (381 mg, 1.003 mmol) were added. The reactionmixture immediately became yellow and was stirred at room temperaturefor 15 minutes. 6-[(2,6-dimethylphenyl)oxy]-3-pyridinamine (Intermediate42, 223.8 mg) was added and the reaction mixture was warmed to 60° C.After 4 hours the solvent, was evaporated under vacuum using the Genevacaffording a dark brown oil. This residue was purified by silica gelchromatography (Biotage instrument, 25 g SNAP Silica column) elutingwith Cyclohexane/EtOAc from 3:1 Cyclohexane/EtOAc to 1:1Cyclohexane/EtOAc in 10 CV; then 1:1 Cyclohexane/EtOAc for 5 CV. Thecollected fractions afforded the title compound as a pale orange oil(282 mg).

¹H NMR (400 MHz, DMSO-d₆) d ppm 10.03 (1H, br. s), 8.22 (1H, d), 8.07(1H, dd), 7.02-7.19 (4H, m), 6.97 (1H, dd), 4.03-4.18 (1H, m), 2.04 (6H,s), 1.40 (9H, s), 1.27 (3H, d); UPLC_B: 0.89 min, 386 [M+H]+.

Intermediate 44N¹-{6-[(2,6-dimethylphenyl)oxy]-3-pyridinyl}-D-alaninamide

In a 50 mL round-bottomed flask 1,1-dimethylethyl[(1R)-2-({6-[(2,6-dimethylphenyl)oxy]-3-pyridinyl}amino)-1-methyl-2-oxoethyl]carbamate(Intermediate 43, 282 mg) was dissolved in dichloromethane (2 mL) togive a yellow solution. Trifluoroacetic acid (2 mL, 26.0 mmol) wasadded. The reaction mixture was stirred at room temperature. After 20min, the solvent was evaporated under vacuum affording a yellow oil thatwas charged on a 5 g SCX cartridge. It was then flushed with 25 mL ofMeOH followed by 25 mL of 2M solution of ammonia in MeOH. The ammoniaeluate was evaporated under vacuum to afford the title compound as ayellow oil which solidified (173.8 mg).

¹H NMR (400 MHz, CDCl₃) d ppm 9.51 (1H, br. s), 8.23-8.12 (2H, m),7.17-7.01 (3H, m), 6.85-6.75 (1H, m), 3.64 (1H, q), 2.13 (6H, s), 1.83(2H, br. s), 1.44 (3H, d); UPLC_B: 0.70 mins, 286 [M+H]+.

Intermediate 45 2-[(2-ethylphenyl)oxy]-5-nitropyridine

In a 20 mL microwave vial 2-chloro-5-nitropyridine (500 mg, 3.15 mmol)was dissolved in N,N-dimethylformamide (10 mL) to give a light brownsolution. 2-ethylphenol (0.378 mL, 3.15 mmol) and K₂CO₃ (1308 mg, 9.46mmol) were added. The reaction vessel was sealed and heated in BiotageInitiator at 110° C. for 1 hour. After cooling the reaction wascomplete. The reaction mixture was quenched with 10 mL of water anddiluted with 10 mL of Et₂O. Phases were separated through a separatingfunnel. The organic phase was washed with 3×10 mL of water, 10 mL ofbrine, dried over sodium sulphate, filtered and evaporated under vacuumto give the title compound as a pale orange oil. (623 mg).

¹H NMR (400 MHz, CDCl₃) δ ppm 9.06 (1H, d), 8.49 (1H, dd), 7.40-7.34(1H, m), 7.33-7.28 (2H, m), 7.10-7.05 (1H, m), 7.04 (1H, d), 2.55 (2H,q), 1.18 (3H, t); UPLC_B: 0.95 min, 245 [M+H]+.

Intermediate 46 6-[(2-ethylphenyl)oxy]-3-pyridinamine

In a 50 mL round-bottomed flask 2-[(2-ethylphenyl)oxy]-5-nitropyridine(Intermediate 45, 623 mg) was dissolved in ethanol (10 mL) to give apale orange solution. Palladium on carbon (244 mg, 0.230 mmol) andhydrazine hydrate (0.442 mL, 4.59 mmol) were added. The reaction mixturewas stirred at 90° C. After 3 hours the reaction was completed. Thereaction mixture was filtered and the organic phase was evaporated undervacuum affording 1.1408 g of a dark orange solid that was charged on a10 g SCX cartridge. It was then flushed with 200 mL of ethanol followedby 50 mL of 2M solution of ammonia in MeOH. The ammonia eluate wasevaporated under vacuum to afford the title compound as dark orangesolid (456.1 mg).

¹H NMR (400 MHz, CDCl₃) δ ppm 7.71 (d, 1H), 7.26-7.31 (m, 1H), 7.16-7.22(m, 1H), 7.10-7.15 (m, 1H), 7.07 (dd, 1H), 6.96 (dd, 1H), 6.70 (d, 1H),3.52 (br. s., 2H), 2.64 (q, 2H), 1.20 (t, 3H); UPLC_B: 0.75 mins, 215[M+H]+.

Intermediate 471,1-dimethylethyl[(1R)-2-({6-[(2-ethylphenyl)oxy]-3-pyridinyl}amino)-1-methyl-2-oxoethyl]carbamate

In a 8 mL vial N-{[(1,1-dimethylethyl)oxy]carbonyl}-D-alanine (193 mg,1.022 mmol) was dissolved in N,N-dimethylformamide (4 mL) to give acolorless solution. N-ethyl-N-(1-methylethyl)-2-propanamine (0.223 mL,1.277 mmol) andN-[(1H-1,2,3-benzotriazol-1-yloxy)(dimethylamino)methylidene]-N-methylmethanaminiumtetrafluoroborate (328 mg, 1.022 mmol) were added. The reaction mixturewas stirred at room temperature for 15 min.6-[(2-ethylphenyl)oxy]-3-pyridinamine (Intermediate 46, 228 mg) wasadded and the reaction mixture was warmed to 60° C. for 32 hours. Thesolvent was evaporated under vacuum using the Genevac affording a darkbrown oil, which was purified by silica gel chromatography (Biotagesystem, 25 g SNAP column) eluting with a gradient Cyclohexane/EtOAc from3:1 C to 1:1 in 10 CV; then 1:1 for 5V. The collected fractions affordedthe title compound (251.1 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 8.70 (1H, br. s), 8.17 (1H, d), 8.05 (1H,dd), 7.31 (1H, dd), 7.14-7.27 (2H, m), 7.02 (1H, dd), 6.82 (1H, d), 5.11(1H, d), 4.22-4.52 (1H, m), 2.60 (2H, q), 1.47 (9H, s), 1.44-1.46 (3H,m), 1.18 (3H, t); UPLC_B: 0.90 min, 386 [M+H]+.

Intermediate 48 N¹-{6-[(2-ethylphenyl)oxy]-3-pyridinyl}-D-alaninamide

In a 50 mL round-bottomed flask 1,1-dimethylethyl[(1R)-2-({6-[(2-ethylphenyl)oxy]-3-pyridinyl}amino)-1-methyl-2-oxoethyl]carbamate(Intermediate 47, 251.1 mg) was dissolved in dichloromethane (2 mL) togive a pale orange solution. Trifluoroacetic acid (2 mL, 26.0 mmol) wasadded. The reaction mixture was stirred at room temperature. After 20minutes the solvent was evaporated under vacuum affording a yellow oilwhich was charged on a 5 g SCX cartridge and flushed with 25 mL of MeOHfollowed by 25 mL of 2M solution of ammonia in MeOH. The ammonia eluatewas evaporated under vacuum to afford the title compound as a yellow oilwhich solidified (170.0 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 9.58 (1H, br. s), 8.24 (1H, d), 8.20 (1H,dd), 7.30-7.35 (1H, m), 7.15-7.27 (2H, m), 7.03 (1H, dd), 6.85 (1H, d),3.69 (1H, q), 2.61 (2H, q), 2.31 (2H, br. s), 1.46 (3H, d), 1.20 (3H,t); UPLC_B: 0.71 mins, 286 [M+H]+.

Intermediate 49 2-{[4-methyl-3-(methyloxy)phenyl]oxy}-5-nitropyridine

To a solution of 4-methyl-3-(methyloxy)phenol (Intermediate 18, 400 mg)in dry N,N-dimethylformamide (15 mL), potassium carbonate (1200 mg, 8.69mmol) and then 2-chloro-5-nitropyridine (551 mg, 3.47 mmol) were addedand the reaction mixture was stirred for 2 hours at 115° C. The reactionwas quenched with water (10 mL), diluted with brine (20 mL) andextracted with ethyl acetate (3 times 30 mL). The organic layer waswashed with ice cold brine (2 times 30 mL), dried over sodium sulphate,filtered and evaporated. The residue was purified by silica gelchromatography (Biotage system, 100 g SNAP column) with a gradientcyclohexane/ethyl acetate from 10/0 to 8/2. Evaporation afforded the totitle compound as a light yellow oil (570 mg).

¹H-NMR (400 MHz, DMSO-d₆) δ ppm: 2.16 (3H, s), 3.76 (3H, s), 6.68-6.73(1H, m), 6.83-6.86 (1H, m), 7.24-7.18 (2H, m), 8.64-8.58 (1H, m),9.08-9.02 (1H, m); UPLC_B: 0.93 min, 261 [M+H]+.

Intermediate 50 6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinamine

To a solution of 2-{[4-methyl-3-(methyloxy)phenyl]oxy}-5-nitropyridine(Intermediate 49, 568 mg) in tetrahydrofuran (25 mL)/water (12.50 mL),iron (609 mg, 10.91 mmol) and then ammonium chloride (584 mg, 10.91mmol) were added and the reaction mixture was stirred for 8 hours atroom temperature. The catalyst was filtered off and the solution wasdiluted with an aqueous saturated solution of Na2CO3 (5 mL) andextracted with ethyl acetate (2 times 40 mL). Combined organic layerswere dried over sodium sulphate, filtered and evaporated and the residuewas purified by silica gel chromatography (Biotage system with a 50 gSNAP column) using a as eluent a gradient cyclohexane/ethyl acetate from8/2 to 1/1. Evaporation afforded the title compound as light yellow oil(465 mg).

¹H-NMR (400 MHz, DMSO-d₆) δ ppm: 7.54 (1H, d), 7.06 (2H, ddd), 6.72 (1H,d), 6.59 (1H, d), 6.38 (1H, dd), 5.07 (2H, s), 3.73 (3H, s), 2.10 (3H,s); UPLC_B: 0.72 min, 231 [M+H]+.

Intermediate 511,1-dimethylethyl{(1R)-1-methyl-2-[(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)amino]-2-oxoethyl}carbamate

To a solution of N-{[(1,1-dimethylethyl)oxy]carbonyl}-D-alanine (181 mg,0.955 mmol) in dry N,N-dimethylformamide (3 mL), DIPEA (0.303 mL, 1.737mmol) and then TBTU (335 mg, 1.042 mmol) were added and the reactionmixture was stirred for 15 minutes at room temperature.6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinamine (Intermediate 50,200 mg) was then added and the reaction mixture was stirred for 1 hourat the same temperature. The reaction was quenched with water (2 mL),diluted with brine (10 mL) and extracted with ethyl acetate (2×20 mL).The organic layer was dried over sodium sulphate, filtered andevaporated and the residue was purified by silica gel chromatography(Biotage system, 10 g SNAP column) using a gradient cHex/EtOAc as eluentfrom 100/0 to 70/30. This afforded the title compound (350 mg)

¹H NMR (400 MHz, DMSO-d₆) d ppm 10.09 (1H, br. s), 8.38-8.29 (1H, m),8.09-8.03 (1H, m), 7.12 (2H, d), 6.97 (1H, d), 6.70 (1H, d), 6.57-6.51(1H, m), 4.16-4.04 (1H, m), 3.74 (3H, s), 2.13 (3H, s), 1.39 (9H, s),1.26 (3H, d); UPLC_B: 0.87 min, 402 [M+H]+

Intermediate 52N¹-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)-D-alaninamide

To a solution of 1,1-dimethylethyl{(1R)-1-methyl-2-[(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)amino]-2-oxoethyl}carbamate(Intermediate 51, 350 mg) in dry dichloromethane (7.5 mL), TFA (2.5 mL,32.4 mmol) was slowly added and the reaction mixture was stirred for 1.5hours at room temperature. The solvent and the excess of TFA wereevaporated and the residue was purified with an SCX cartridge (10 g) toafford the title compound as a colourless oil (258 mg).

¹H NMR (400 MHz, DMSO-d₆) d ppm 8.40 (1H, d), 8.11 (1H, dd), 7.13 (1H,d), 6.96 (1H, d), 6.70 (1H, d), 6.54 (1H, dd), 3.75 (3H, s), 3.44 (1H,q), 3.33 (2H, br. s), 2.13 (3H, s), 1.22 (3H, d); UPLCB: 0.70 min, 302[M+H]+

Intermediate 53 2-{[2-methyl-5-(methyloxy)phenyl]oxy}-5-nitropyridine

To a solution of 2-methyl-5-(methyloxy)phenol (Intermediate 12, 2 g) and2-chloro-5-nitropyridine (2.1 g, 13.2 mmol) in DMF (50 mL) was addedpotassium carbonate (2.76 g, 20 mmol) and the reaction mixture wasstirred at 100° C. overnight. The mixture was evaporated under vacuumand water (100 mL) was added. It was extracted with ethyl acetate (3times 100 mL) and the combined organic layers were dried over sodiumsulphate and evaporated to afford a brown oil, which was purified bycolumn chromatography on silica gel (mobile phase: EtOAc/PE=1/50−1/20)to give the title compound (1.5 g).

MS_(—)2 (ESI): 261 [M+H]+

Intermediate 54 6-{[2-methyl-5-(methyloxy)phenyl]oxy}-3-pyridinamine

To a solution of 2-{[2-methyl-5-(methyloxy)phenyl]oxy}-5-nitropyridine(Intermediate 53, 1.5 g) in ethanol (100 mL) was added Pd/C (5%, 200 mg)and the mixture was stirred at room temperature under H₂ atmosphereovernight. The mixture was filtered through a pad of celite and thefiltrate was evaporated to afford a yellow oil, which was purified bycolumn chromatography on silica gel (mobile phase: EtOAc/PE=1/5−1/2).This afforded the title compound (850 mg).

¹HNMR (400 MHz, CDCl₃): δ 7.70 (1H, s), 7.12 (1H, d), 7.05-7.08 (1H, d),6.69 (1H, d), 6.63-6.66 (1H, d), 6.54 (1H, s), 3.74 (3H, s), 3.45 (2H,s), 2.12 (3H, s); MS_(—)2 (ESI): 231 [M+H]+

Intermediate 551,1-dimethylethyl{(1R)-1-methyl-2-[(6-{[2-methyl-5-(methyloxy)phenyl]oxy}-3-pyridinyl)amino]-2-oxoethyl}carbamate

The title compound was made in a similar fashion to the preparation ofIntermediate 15 replacing 4-{[2-methyl-5-(methyloxy)phenyl]oxy}anilinewith 6-{[2-methyl-5-(methyloxy)phenyl]oxy}-3-pyridinamine (Intermediate54, 90 mg) to afford the title compound (152 mg).

¹H NMR (400 MHz, methanol-d₄): δ ppm 8.32 (1H, d), 8.05 (1H, d), 7.18(1H, d), 6.86 (1H, d), 6.76 (1H, d), 6.61 (1H, d), 4.25 (1H, m), 3.13(3H, s), 2.08 (3H, s), 1.46 (9H, s), 1.41 (3H, d); UPLC_B: 0.85 min, 402[M+H]+

Intermediate 56N¹-(6-{[2-methyl-5-(methyloxy)phenyl]oxy}-3-pyridinyl)-D-alaninamide

The title compound was made in a similar fashion to the preparation ofIntermediate 16 replacing{(1R)-1-methyl-2-[(4-{[2-methyl-5-(methyloxy)phenyl]oxy}phenyl)amino]-2-oxoethyl}carbamatewith1,1-dimethylethyl{(1R)-1-methyl-2-[(6-{[2-methyl-5-(methyloxy)phenyl]oxy}-3-pyridinyl)amino]-2-oxoethyl}carbamate(Intermediate 55, 150 mg) to afford the title compound (120 mg).

¹H NMR (400 MHz, methanol-d₄): δ ppm 8.32 (1H, d), 8.05 (1H, dd), 7.17(1H, d), 6.83 (1H, d), 6.73 (1H, d), 6.58 (1H, d), 3.74 (3H, s),3.63-3.50 (1H, m), 2.06 (3H, s), 1.36 (3H, d); UPLC_B: 0.66 min, 302[M+H]+

Intermediate 57 2-methyl-1-(methyloxy)-3-nitrobenzene

To a solution of 2-methyl-3-nitrophenol (15.3 g, 100 mmol) in DMF (150mL) was added sodium hydride (60% in mineral oil, 2.6 g, 110 mmol) at 0°C. and the mixture was stirred for 30 minutes at room temperature.Methyl iodide (28.4 g, 200 mmol) was added and the mixture was heated to80° C. for 5 hours. Water (100 mL) was added and the mixture wasextracted with ethyl acetate (3 times 100 mL). The combined ethylacetate phases were dried over sodium sulphate and concentrated undervacuum to give a residue, which was purified by column chromatography onsilica gel (PE:EtOAc=5:1). Evaporation afforded the title compound as ayellow solid (14.4 g).

Intermediate 58 2-methyl-3-(methyloxy)aniline

To a solution of 2-methyl-1-(methyloxy)-3-nitrobenzene (Intermediate 57,1.67 g) in methanol (50 mL) was added Pd/C (10%, 50 mg) and the reactionmixture was stirred under H2 atmosphere for 1 hour. The mixture wasfiltered through a pad of celite. Evaporation afforded the titlecompound as a solid (1.31 g).

Intermediate 59 2-methyl-3-(methyloxy)phenol

To a solution of 2-methyl-3-(methyloxy)aniline (Intermediate 58, 1.31 g)in H2SO4 (6 M, 100 mL) was added portionwise NaNO2 (794 mg, 11 mmol) at0° C. The mixture was stirred for another 2 hours at 40° C. and water(100 mL) was added. The resulting mixture was extracted with ethylacetate (3 times 100 mL) and the combined ethyl acetate phases weredried and evaporated. The residue was purified by silica gel columnchromatography (PE: EtOAc=5:1) to afford the title compound as a solid(569 mg).

Intermediate 60 2-{[2-methyl-3-(methyloxy)phenyl]oxy}-5-nitropyridine

To a solution of 2-methyl-3-(methyloxy)phenol (Intermediate 59, 1.3 g)in DMF (50 mL) was added sodium hydride (60% in mineral oil, 480 mg,0.012 molat 0° C. and the mixture was stirred for 30 minutes.2-Chloro-5-nitropyridine (1.9 g, 0.012 mol, Aldrich) was added and themixture was heated at 60° C. for 3 hours. The mixture was poured intoH2O (100 mL) and extracted with ethyl acetate (4 times 100 mL). Thecombined ethyl acetate phases were dried over sodium sulphate andconcentrated under vacuum to give a residue which was purified by columnchromatography on silica gel (PE: EtOAc=10:1) to afford the titlecompound (2.3 g) as a liquid.

MS_(—)2 (ESI): 261 [M+H]+.

Intermediate 61 6-{[2-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinaminehydrochloride salt

To a solution of 2-{[2-methyl-3-(methyloxy)phenyl]oxy}-5-nitropyridine(Intermediate 60, 2.3 g) in methanol (30 mL) was added Pd/C (10%, 0.3 g)and H2 was bubbled into the mixture for 2 hours at room temperature. Thereaction mixture was filtered through a pad of Celite. The filtrate wasbubbled into HCl gas. The resulting mixture was concentrated to affordthe title compound as a white solid (2 g).

¹HNMR (DMSO-d₆): δ ppm 10.0-8.5 (3H, m), 8.03-8.02 (1H, s), 7.73-7.71(1H, d), 7.22-7.18 (1H, t), 7.05-7.02 (1H, d), 6.87-6.85 (1H, d),6.65-6.63 (1H, d), 3.82 (3H, s), 1.90 (3H, s);

MS_(—)2 (ESI): 231 [M−(HCl)+H]+.

Intermediate 621,1-dimethylethyl{(1R)-1-methyl-2-[(6-{[2-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)amino]-2-oxoethyl}carbamate

The title compound (307 mg) was made in a similar fashion to thepreparation of Intermediate 25 replacing6-{[3-(1-methylethyl)phenyl]oxy}-3-pyridinamine with6-{[2-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinamine (Intermediate 61,201 mg).

¹H NMR (400 MHz, methanol-d₄): δ ppm 8.75 (1H, d), 8.44 (1H, dd),7.93-8.11 (1H, m), 7.53 (1H, dd), 7.19 (1H, t), 6.83 (2H, t), 6.62 (1H,d), 4.09-4.33 (1H, m), 3.87 (3H, s), 2.02 (3H, s), 1.44-1.51 (9H, m),1.42 (3H, d); UPLC_B: 0.86 min, 402 [M+H]+

Intermediate 63N¹-(6-{[2-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)-D-alaninamide

The title compound (268 mg) was made in a similar fashion to thepreparation of Intermediate 26 replacing1,1-dimethylethyl{(1R)-1-methyl-2-[(6-{[3-(1-methylethyl)phenyl]oxy}-3-pyridinyl)amino]-2-oxoethyl}carbamatewith1,1-dimethylethyl{(1R)-1-methyl-2-[(6-{[2-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)amino]-2-oxoethyl}carbamate(Intermediate 62, 304 mg). Instead of being stirred at 0° C. for 3hours, the reaction was stirred at 0° C. for 1 hour and at roomtemperature for 2 hours.

¹H NMR (400 MHz, methanol-d₄): δ ppm 8.33 (1H, d), 8.03 (1H, dd), 7.18(1H, t), 6.81 (2H, t), 6.60 (1H, d), 3.85 (3H, s), 3.66 (1H, q), 2.00(3H, s), 1.40 (3H, d); UPLC_B: 0.67 min, 302 [M+H]+

Intermediate 641,1-dimethylethyl((1R)-1-{[(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)amino]carbonyl}propyl)carbamate

To a solution of(2R)-2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)butanoic acid (106 mg,0.521 mmol) in dry N,N-dimethylformamide (2 mL) DIPEA (0.152 mL, 0.869mmol) and then TBTU (181 mg, 0.565 mmol) were added and the reactionmixture was stirred for 15 minutes at room temperature.6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinamine (Intermediate 50,100 mg) was then added and the reaction mixture was stirred overnight atthe same temperature. The reaction was quenched with water (1 mL),diluted with brine (1 mL) and extracted with ethyl acetate (3 times 5mL). The organic layer was dried over sodium sulphate, filtered andevaporated and the residue was purified by silica gel chromatography(Biotage system, 10 g SNAP column) using as eluent a gradientcyclohexane/ethyl acetate from 100/0 to 70/30 to afford the titlecompound as a white solid (180 mg).

¹H NMR (400 MHz, DMSO-d₆) δ ppm: 10.13 (1H, br. s), 8.31-8.37 (1H, m),8.02-8.10 (1H, m), 7.09-7.16 (1H, m), 7.01-7.08 (1H, m), 6.96 (1H, d),6.70 (1H, d), 6.51-6.58 (1H, m), 3.91-4.03 (1H, m), 3.75 (3H, s), 2.13(3H, s), 1.50-1.76 (2H, m), 1.39 (9H, s), 0.90 (3H, t); UPLC_B: 0.91min, 416 [M+H]+.

Intermediate 65(2R)-2-amino-N-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)butanamide

To a solution of 1,1-dimethylethyl((1R)-1-{[(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)amino]carbonyl}propyl)carbamate(Intermediate 64, 175 mg) in dry dichloromethane (DCM) (6 mL) TFA (2 mL,26.0 mmol) was slowly added and the reaction mixture was stirred for 1 hat room temperature. The solvent and the excess of TFA were evaporatedand the residue was purified by SCX cartridge (5 g) to afford the titlecompound as a colourless solid (122 mg).

¹H-NMR (400 MHz, DMSO-d₆) δ ppm: 8.36-8.42 (1H, m), 8.11 (1H, dd), 7.12(1H, d), 6.95 (1H, d), 6.67-6.73 (1H, m), 6.54 (1H, dd), 3.75 (3H, s),3.24 (1H, m), 2.13 (3H, s), 1.59-1.73 (1H, m), 1.42-1.56 (1H, m), 0.90(3H, t); UPLC_B: 0.74 min, 316 [M+H]+.

Intermediate 65b 2HCl Salt of Intermediate 65(2R)-2-amino-N-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)butanamide.2HCl

6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinamine (Intermediate 50,500 g), (2R)-2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)butanoic acid(530 g) and Et₃N (905 mL) are mixed together in ethyl acetate (2 L) andstirred at 0° C. until complete dissolution. ®T3P (2.15 L) was addeddropwise in 30 minutes maintaining the temperature at −0° C. Ethylacetate (500 mL) was added for washing the line. Work-up: 10% w/w sodiumcarbonate aqueous solution (2.5 L) was added and the mixture was stirredfor 20 minutes. Then water (1.5 L) and ethyl acetate (1 L) were addedand the two phases separated. The organic layer was washed with 10% w/wsodium carbonate aqueous solution (2.5 L), stirring the mixture for 10minutes before separation of phases, then with 28% malic acid aqueoussolution (2.5 L) and finally with 20% NaCl aqueous solution (2.5 L). Theorganic solution was concentrated to the lowest volume (<2 L),acetonitrile (5 L) was added, the solution was concentrated to thelowest volume (<2 L) and acetonitrile was added up to 12.5 L (it is asolution of Intermediate 64 in acetonitrile). To this solution, 5-6N HClsolution in Isopropanol (2.5 L) was added at 20° C. and the resultingreaction mixture was stirred at 45° C. for 1.5 hours. The obtainedsuspension was cooled to 20° C., stirred for 1 hour and then filtered.The collected solid was washed with 5/1 acetonitrile/Isopropanol (3×1.5L), then dried under vacuum at 40° C., until constant weight, obtainingthe title compound (817 g).

¹H-NMR (400 MHz, DMSO-d₆): δ ppm 11.13 (1H, s), 8.30-8.50 (m, 4H), 8.07(1H, dd), 7.10 (1H, d), 6.98 (1H, d), 6.69 (1H, d), 6.53 (1H, dd), 3.99(1H, m), 3.72 (3H, s), 2.10 (3H, s), 1.80-1.95 (2H m), 0.92 (3H, t).

A generic Ion Chromatography method was used in order to determine theamount of Chloride. Method Conditions: Equipment Dionex ICS2000, Columntype Dionex AS18 2 mm×250 mm; Mobile Phase KOH 41 mM; Flow rate 0.47mL/min; Conductimetric Detection. Result: Chloride 17.5% w/w. From thisresult Intermediate 65b was confirmed being a di-hydrochloride salt.

Intermediate 661,1-dimethylethyl((1S)-1-{[(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)amino]carbonyl}propyl)carbamate

To a solution of(2S)-2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)butanoic acid (66.2 mg,0.326 mmol) in dry N,N-dimethylformamide (1 mL), DIPEA (0.095 mL, 0.543mmol) and then TBTU (112 mg, 0.347 mmol) were added and the reactionmixture was stirred for 15 minutes at room temperature.6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinamine (Intermediate 50,50 mg) was then added and the reaction mixture was stirred at roomtemperature overnight. The mixture was diluted with ethyl acetate (10mL) and washed with brine (3 times 8 mL). The organic phase wasseparated, dried over sodium sulphate, filtered and evaporated. Theresidue was purified by flash chromatography on silica gel using acolumn SNAP 10 g and cyclohexane/ethyl acetate from 100/0 to 60/40 aseluent. This afforded the title compound as a white solid (73 mg).

¹H NMR (400 MHz, DMSO): δ ppm 10.13 (1H, s), 8.33 (1H, d), 8.06 (1H,dd), 7.11 (1H, d), 7.05 (1H, d), 6.95 (1H, d), 6.69 (1H, d), 6.53 (1H,dd), 4.00-3.91 (1H, m), 3.73 (3H, s), 2.13 (3H, s), 1.74-1.51 (2H, m),1.38 (9H, s), 0.89 (3H, t); UPLC_B: 0.92 min, 414 [M−H]⁻

Intermediate 67(2S)-2-amino-N-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)butanamide

To a solution of 1,1-dimethylethyl((1S)-1-{[(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)amino]carbonyl}propyl)carbamate(Intermediate 66, 70 mg) in dichlorometane (2.5 mL) cooled down to 0° C.TFA (0.779 mL, 10.11 mmol) was added dropwise. The reaction mixture wasstirred at 0° C. for 1.5 hours and then evaporated. The residue wasdiluted with dichloromethane (10 mL) and neutralized with a saturatedaqueous solution of NaHCO₃ (15 mL). The organic phase was separated,dried over sodium sulphate, filtered and evaporated to afford the titlecompound as a yellow pale oil (53.1 mg).

¹H NMR (400 MHz, DMSO): δ ppm 8.39 (1H, d), 8.11 (1H, dd), 7.12 (1H, d),6.95 (1H, d), 6.70 (1H, d), 6.54 (1H, dd), 3.74 (3H, s), 3.52-3.21 (1H,m), 2.12 (3H, s), 1.71-1.44 (2H, m), 0.89 (3H, t). UPLC_B: 0.75 min, 314[M−H]⁻

Intermediate 68 (2R)-2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)butanoicacid

To a solution of (2R)-2-aminobutyric acid 5 (1.95 g, 18.91 mmol) in 19mL of 1 M aqueous sodium hydroxide and 13 mL of methanol was addedBoc-anhydride (4.95 g, 22.69 mmol) at 0° C. The reaction mixture waswarmed to room temperature and stirred for 12 hours. After most of themethanol was evaporated, the solution was acidified to pH 2 with 1 M HCland extracted with ethyl acetate (3 times 60 mL). The organic extractswere combined and washed with brine (2 times 12 mL). Evaporation of thesolvent afforded the title compound (3.48 g).

¹H-NMR (400 MHz, DMSO-d6): δ ppm 12.35 (1H, s), 7.02 (1H, d), 3.71-4.07(1H, m), 1.47-1.79 (2H, m), 1.38 (9H, s), 0.88 (3H, t); UPLC: 0.60 min,204 [M+H]]+.

Intermediate 691,1-dimethylethyl{(1R)-1-methyl-2-[(6-{[3-(1-methylethyl)phenyl]oxy}-3-pyridinyl)amino]-2-oxoethyl}carbamate

To a solution of(2R)-2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)butanoic acid(Intermediate 68, 875 mg) in dry N,N-dimethylformamide (50 mL), DIPEA(0.935 mL, 5.36 mmol, 1.5 equiv) and then HATU (1.629 g, 4.28 mmol, 1.2equiv) were added and the reaction mixture was stirred for 15 minutes atroom temperature under argon.

Then (4-{[3-(methyloxy)phenyl]oxy}phenyl)amine4-{[3-(methyloxy)phenyl]oxy}aniline (Intermediate 24, 815 mg) was addedand the reaction mixture was stirred at 63° C. under argon. The reactionwas left under heating 17 hours. After evaporation, the residue obtainedwas purified by silica gel chromatography (Companion system, 120 gcartridge) with a gradient cyclohexane/ethyl acetate from 100/0 to70/30. The title compound was obtained as a yellow powder (1.282 g)

¹H NMR (400 MHz, methanol-d₄): δ ppm 8.37 (1H, d), 8.10 (1H, dd), 7.34(1H, t), 7.13 (1H, d), 6.98 (1H, t), 6.92 (2H, m), 4.11 (1H, t),2.94-2.89 (1H, m), 2.02-1.77 (1H, m), 1.75-1.72 (1H, m), 1.48 (9H, s),1.27 (6H, d), 1.04 (3H, t); UPLC: 0.91 min, 414 [M+1]+

Intermediate 70(2R)-2-amino-N-(6-{[3-(1-methylethyl)phenyl]oxy}-3-pyridinyl)butanamide

1,1-dimethylethyl((1R)-1-{[(6-{[3-(1-methylethyl)phenyl]oxy}-3-pyridinyl)amino]carbonyl}propyl)carbamate(Intermediate 69, 1.28 g) was dissolved in 18 mL of dry dichloromethane.To this solution at 0° C. under argon was added dropwise 30 equivalentsof TFA (7.15 mL, 93 mmol). The reaction was stirred during 3 hours at 0°C. The reaction mixture was evaporated. The residue obtained waspurified by SCX on a 50 g cartridge. The cartridge was washed with 3 CVof methanol, then the compound was adsorbed on the cartridge, washedwith 5 CV of methanol and desorbed with 2 CV of methanolic ammonia (1N).After evaporation of the volatiles, the title compound was obtained (932mg).

¹H NMR (400 MHz, methanol-d₄): δ ppm 8.36 (1H, d), 8.08 (1H, dd), 7.30(1H, t), 7.08 (1H, d), 6.98-6.78 (3H, m), 3.39 (1H, t), 2.94-2.84 (1H,m), 2.81 (2H, s), 1.87-1.74 (1H, m), 1.73-1.59 (1H, m), 1.25 (6H, d),1.00 (3H, t); UPLC: 0.60 min, 314 [M+1]+

Intermediate 71 1,1-dimethylethyl{1,1-dimethyl-2-[(4-{[3-(methyloxy)phenyl]oxy}phenyl)amino]-2-oxoethyl}carbamate

To a solution of N-{[(1,1-dimethylethyl)oxy]carbonyl}-2-methylalanine(1.7 g, 8.36 mmol) in dry N,N-dimethylformamide (35 mL), DIPEA (2.434mL, 13.94 mmol) and then TBTU (2.80 g, 8.71 mmol) were added and thereaction mixture was stirred for 15 minutes at room temperature.(4-{[3-(methyloxy)phenyl]oxy}phenyl)amine (1.5 g, 6.97 mmol) was thenadded and the reaction mixture was stirred overnight at the sametemperature. The reaction was quenched with brine (100 mL) and extractedwith ethyl acetate (twice 150 mL). The organic layer was washed with icecold brine (3 times 100 mL), dried over sodium sulphate, filtered andevaporated. The residue was purified by silica gel chromatography(Biotage system, 100 g SNAP column) using as eluent a gradient andcyclohexane/ethyl acetate from 100/0 to 70/30 to afford the titlecompound as a white solid (1.90 g).

¹H NMR (400 MHz, DMSO-d₆) d ppm 9.47 (1H, br. s), 7.50-7.76 (2H, m),7.24 (1H, t), 6.97 (3H, d), 6.67 (1H, dd), 6.44-6.55 (2H, m), 3.72 (3H,s), 1.25-1.47 (15H, m); UPLC_B: 0.91 min, 401 [M+1]+.

Intermediate 722-methyl-N¹-(4-{[3-(methyloxy)phenyl]oxy}phenyl)alaninamide

To a solution of 1,1-dimethylethyl{1,1-dimethyl-2-[(4-{[3-(methyloxy)phenyl]oxy}phenyl)amino]-2-oxoethyl}carbamate(Intermediate 71, 1.89 g) in dry dichloromethane (60 mL) at 0° C. TFA(20 mL, 260 mmol) was added dropwise and the reaction mixture wasstirred for 2 hours at the same temperature. The solvent and the excessof TFA were evaporated and the residue was purified by SCX cartridge (50g) to afford the title compound as a light brown oil (1.34 g).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 7.73-7.64 (2H, m), 7.25 (1H, t),6.95-7.03 (2H, m), 6.64-6.70 (1H, m), 6.45-6.55 (2H, m), 3.72 (3H, s),1.28 (6H, s); UPLC_B: 0.79 min, 301 [M+1]+

Intermediate 73 2,3-dimethylphenyl 4-nitrophenyl ether

In a microwave vial 1-fluoro-4-nitrobenzene (500 mg, 3.54 mmol) wasdissolved in N,N-Dimethylformamide (10 mL) to give a pale yellowsolution. Potassium carbonate (1469 mg, 10.63 mmol) and2,3-dimethylphenol (433 mg, 3.54 mmol) were added. The reaction vesselwas sealed and heated in Biotage Initiator at 100° C. for 1 hour. Aftercooling the reaction was diluted with 25 mL of Et₂O. The organic phasewas washed with 3 times 25 mL of water, 10 mL of saturated brine, driedover sodium sulphate, filtered and evaporated under vacuum to afford thetitle compound as a yellow solid (865.1 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 8.25-8.14 (2H, m), 7.17 (1H, t),7.13-7.08 (1H, m), 6.96-6.83 (3H, m), 2.36 (3H, s), 2.10 (3H, s); UPLC:0.90 min, 244 [M+H]+.

Intermediate 74 4-[(2,3-dimethylphenyl)oxy]aniline

2,3-dimethylphenyl 4-nitrophenyl ether (Intermediate 73, 865 mg) wasdissolved in ethanol (10 mL) to give a pale yellow solution. Hydrazinehydrate 50% (0.698 mL, 7.1 mmol) and Pd/C (37.8 mg, 0.36 mmol) wereadded. The reaction mixture was stirred at 90° C. for 1 hour. Thereaction mixture was filtered and the organic phase was evaporated undervacuum to afford the title compound as a pale yellow oil (796 mg).

¹H-NMR (400 MHz, CDCl₃): δ ppm 7.07-6.96 (1H, m), 6.90 (1H, d),6.83-6.73 (2H, m), 6.72-6.61 (3H, m), 2.33 (3H, s), 2.22 (3H, s); UPLC:0.60 min, 214 [M+H]+.

Intermediate 751,1-dimethylethyl[2-({4-[(2,3-dimethylphenyl)oxy]phenyl}amino)-1,1-dimethyl-2-oxoethyl]carbamate

4-[(2,3-dimethylphenyl)oxy]aniline (Intermediate 74, 200 mg) wasdissolved in 5.0 mL of DMF. Then DIPEA (0.246 mL, 1.41 mmol) and HATU(428 mg, 1.13 mmol) were added. After stirring for 15 minutes,N-{[(1,1-dimethylethyl)oxy]carbonyl}-2-methylalanine (229 mg, 1.13 mmol)was added and the reaction mixture was stirred at 40° C. overnight.After removal of the volatiles, the crude was purified by silica gelchromatography eluting with a gradient cHex/EtOAc from 100/0 to 0/100 toafford the title compound (109 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 7.51-7.41 (2H, m), 7.12-6.95 (2H, m),6.93-6.84 (2H, m), 6.82-6.73 (1H, m), 2.35 (3H, s), 2.19 (3H, s), 1.60(3H, s), 1.57 (3H, s), 1.49 (9H, s);

UPLC: 0.83 min, 399 [M+H]+.

Intermediate 76N¹-{4-[(2,3-dimethylphenyl)oxy]phenyl}-2-methylalaninamide

1,1-dimethylethyl[2-({4-[(2,3-dimethylphenyl)oxy]phenyl}amino)-1,1-dimethyl-2-oxoethyl]carbamate(Intermediate 75, 109 mg) was dissolved in 4.0 mL of dichloromethane andthen TFA (1.0 mL) was added. The reaction mixture was stirred at roomtemperature for 1 hour. After the removal of the volatiles, the residuewas charged on a SCX cartridge and eluted with DCM/MeOH/NH3 (2.0 Msolution in MeOH). Evaporation afforded 68 mg of the title compound.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 7.62 (2H, d), 7.17-6.96 (2H, m), 6.84(2H, d), 6.78-6.68 (1H, m), 2.30 (3H, s), 2.11 (3H, s), 1.29 (6H, s);UPLC: 0.57 min, 299 [M+H]+.

Intermediate 77 1,1-dimethylethyl[2-({6-[(2-ethylphenyl)oxy]-3-pyridinyl}amino)-1,1-dimethyl-2-oxoethyl]carbamate

In a 8 mL vial N-{[(1,1-dimethylethyl)oxy]carbonyl}-2-methylalanine (208mg, 1.022 mmol) was dissolved in N,N-dimethylformamide (4 mL) to give acolourless solution. N-ethyl-N-(1-methylethyl)-2-propanamine (0.223 mL,1.277 mmol) andN-[(1H-1,2,3-benzotriazol-1-yloxy)(dimethylamino)methylidene]-N-methylmethanaminiumtetrafluoroborate (328 mg, 1.022 mmol) were added. The reaction mixturewas stirred at room temperature for 15 min.6-[(2-ethylphenyl)oxy]-3-pyridinamine (Intermediate 46, 228 mg) wasadded and the reaction mixture was warmed to 60° C. After 24 hours,additional 150 mg ofN-[(1H-1,2,3-benzotriazol-1-yloxy)(dimethylamino)methylidene]-N-methylmethanaminiumtetrafluoroborate were added. After additional 8 hours, the solvent wasevaporated under vacuum using the Genevac affording a dark brown oilwhich was purified by silica gel chromatography (Biotage system, 25 gSNAP column) with as eluent a gradient Cyclohexane/EtOAc from 3:1 to 1:1in 10 CV, then 1:1 for 5 CV. The collected fractions afforded the titlecompound as a pale orange solid (88.6 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 9.06 (1H, br. s), 8.17-8.10 (2H, m), 7.33(1H, dd), 7.29-7.15 (2H, m), 7.04 (1H, dd), 6.91-6.82 (1H, m), 4.91 (1H,br. s), 2.62 (2H, q), 1.60 (6H, s), 1.47 (9H, s), 1.20 (3H, t); UPLC_B:0.92 min, 400 [M+H]+.

Intermediate 78N¹-{6-[(2-ethylphenyl)oxy]-3-pyridinyl}-2-methylalaninamide

In a 50 mL round-bottomed flask 1,1-dimethylethyl[2-({6-[(2-ethylphenyl)oxy]-3-pyridinyl}amino)-1,1-dimethyl-2-oxoethyl]carbamate(Intermediate 77, 88.6 mg) was dissolved in dichloromethane (2 mL) togive a yellow solution. Trifluoroacetic acid (2 mL, 26.0 mmol) wasadded. The reaction mixture was stirred at room temperature. After 20minutes, the solvent was evaporated under vacuum affording a yellow oilthat was charged on a 5 g SCX cartridge and was then flushed with 25 mLof MeOH followed by 25 mL of 2M solution of ammonia in MeOH. The ammoniaeluate was evaporated under vacuum to afford the title compound as ayellow oil which solidified (67.1 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 9.90 (1H, br. s), 8.18-8.27 (2H, m), 7.33(1H, dd), 7.15-7.28 (2H, m), 7.03 (1H, dd), 6.80-6.90 (1H, m), 2.62 (2H,q), 1.84 (2H, br. s), 1.49 (6H, s), 1.20 (3H, t); UPLC_B: 0.78 min, 300[M+H]+.

Intermediate 79 1,1-dimethylethyl[2-({6-[(2,6-dimethylphenyl)oxy]-3-pyridinyl}amino)-1,1-dimethyl-2-oxoethyl]carbamate

In a 8 mL vial N-{[(1,1-dimethylethyl)oxy]carbonyl}-2-methylalanine (204mg, 1.003 mmol) and N-ethyl-N-(1-methylethyl)-2-propanamine (0.219 mL,1.253 mmol) were dissolved in N,N-Dimethylformamide (4 mL) to give apale yellow solution.N-[(dimethylamino)(3H-[1,2,3]triazolo[4,5-b]pyridin-3-yloxy)methylidene]-N-methylmethanaminiumhexafluorophosphate (381 mg, 1.003 mmol) was added. The reaction mixturebecame bright yellow and was stirred at room temperature for 15 minutes.6-[(2,6-dimethylphenyl)oxy]-3-pyridinamine (Intermediate 42, 223.8 mg)was added and the reaction mixture was warmed at 60° C. After 4 hours,the reaction was complete. The solvent was evaporated under vacuum usingthe Genevac affording a dark brown oil which was purified by silica gelchromatography (Biotage system, 25 g SNAP column) using as eluents agradient Cyclohexane/EtOAc from 3:1 to 1:1 in 10 CV; then 1:1 for 5 CV.The collected fractions afforded the title compound as a pale yellowsolid (202.1 mg).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 9.55 (1H, br. s), 8.15-8.28 (1H, m),7.99-8.11 (1H, m), 7.10-7.18 (2H, m), 7.04-7.10 (1H, m), 6.97-7.05 (1H,m), 6.94 (1H, d), 2.04 (6H, s), 1.38 (15H, br. s); UPLC_B: 0.91 min, 400[M+H]+.

Intermediate 80N¹-{6-[(2,6-dimethylphenyl)oxy]-3-pyridinyl}-2-methylalaninamide

In a 50 mL round-bottomed flask 1,1-dimethylethyl[2-({6-[(2,6-dimethylphenyl)oxy]-3-pyridinyl}amino)-1,1-dimethyl-2-oxoethyl]carbamate(Intermediate 79, 202.1 mg) was dissolved in dichloromethane (2 mL) togive a pale yellow solution. Trifluoroacetic acid (2 mL, 26.0 mmol) wasadded. The reaction mixture was stirred at room temperature for 20minutes. The solvent was evaporated under vacuum to afford a yellow oilwhich was charged on a 5 g SCX cartridge. It was then flushed with 25 mLof MeOH followed by 25 mL of 2M solution of ammonia in MeOH. The ammoniaeluate was evaporated under vacuum to afford the title compound as ayellow oil which solidified (144.4 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 9.87 (1H, br. s), 8.21 (1H, dd), 8.17(1H, d), 7.03-7.16 (3H, m), 6.79 (1H, d), 2.14 (6H, s), 1.85 (2H, br.s), 1.47 (6H, s). UPLC_B: 0.77 min, 300 [M+H]+.

Intermediate 81 N-{[(1,1-dimethylethyl)oxy]carbonyl}-D-valine

To a solution of D-valine (1 g, 8.54 mmol) in tetrahydrofuran (40 mL) asolution of NaOH (0.376 g, 9.39 mmol) in water (10 mL) was addedfollowed by the addition of Boc-Anhydride (2.180 mL, 9.39 mmol). Thereaction mixture was stirred overnight at room temperature. HCl 5% inwater was added while the pH was allowed to reach ˜5-6 and the mixturewas extracted with ethyl acetate (50 mL). Combined organic layers weredried sodium sulphate, filtered and evaporated to afford the titlecompound as a colourless oil (1.85 g).

¹H-NMR (400 MHz, DMSO-d₆) δ ppm: 12.53 (1H, br.s), 6.91-6.80 (1H, m),3.76 (1H, dd), 2.06-1.92 (1H, m), 1.38 (9H, s), 0.86 (6H, t).

Intermediate 821,1-dimethylethyl((1R)-2-methyl-1-{[(4-{[4-methyl-3-(methyloxy)phenyl]oxy}phenyl)amino]carbonyl}propyl)carbamate

To a solution of N-{[(1,1-dimethylethyl)oxy]carbonyl}-D-valine(Intermediate 81, 120 mg) in dry N,N-dimethylformamide (1 mL), DIPEA(0.152 mL, 0.872 mmol) and then TBTU (182 mg, 0.567 mmol) were added andthe reaction mixture was stirred for 5 minutes at room temperature.4-{[4-methyl-3-(methyloxy)phenyl]oxy}aniline (Intermediate 50, 100 mg)was then added and the reaction mixture was stirred for 3 hours at thesame temperature. The reaction was quenched with brine (2 mL) andextracted with ethyl acetate (3 times 3 mL), the organic layer was driedover sodium sulphate, filtered and evaporated. The residue was purifiedby silica gel chromatography (Biotage system, 10 g SNAP column) using aseluent a gradient cyclohexane/ethyl acetate from 100/0 to 80/20 toafford the title compound (113 mg) as a white solid.

UPLC_B: 1.04 min, 429 [M+H]+.

Intermediate 83N¹-(4-{[4-methyl-3-(methyloxy)phenyl]oxy}phenyl)-D-valinamide

To a solution of 1,1-dimethylethyl((1R)-2-methyl-1-{[(4-{[4-methyl-3-(methyloxy)phenyl]oxy}phenyl)amino]carbonyl}propyl)carbamate(Intermediate 82, 110 mg) in dry dichloromethane (3 mL), TFA (1 mL,12.98 mmol) was added and the reaction mixture was stirred for 1 hour atroom temperature. The solvent and the excess of TFA were evaporated andthe residue was purified by SCX cartridge (5 g) to afford the titlecompound as a yellow pale solid (68 mg).

¹H-NMR (400 MHz, DMSO-d₆) δ ppm: 0.89 (dd, 6H), 7.66-7.60 (2H, m),7.12-7.06 (1H, m), 7.00-6.92 (2H, m), 6.67-6.61 (1H, m), 6.42-6.36 (1H,m), 3.74 (3H, s), 3.13-3.07 (1H, m), 2.11 (3H, s), 1.98-1.88 (1H, m);UPLC_B: 0.89 min, 329 [M+H]+.

Intermediate 84 2-{[3-(1-methylethyl)phenyl]oxy}-5-nitropyrimidine

To a solution of 3-(1-methylethyl)phenol (680 mg, 5 mmol, Aldrich) inacetonitrile (50 mL) were added 2-chloro-5-nitropyrimidine (800 mg, 5mmol) and triethylamine (1.01 g, 10 mmol) and the resulting mixture washeated at reflux and stirred for 3 hours. The reaction mixture wasconcentrated under vacuum and water was added to the residue (80 mL). Itwas extracted with ethyl acetate (3 times 50 mL) and the combinedorganic layers were dried over sodium sulphate and evaporated to afforda brown oil, which was purified by silica gel chromatography (mobilephase: ethyl acetate: petroleum ether=0˜20%) to afford the titlecompound (900 mg).

MS_(—)2 (ESI): 260 [M+H]+

Intermediate 85 2-{[3-(1-methylethyl)phenyl]oxy}-5-pyrimidinamine

To a solution of 2-{[3-(1-methylethyl)phenyl]oxy}-5-nitropyrimidine(Intermediate 84, 520 mg) in methanol (50 mL) was added Pd/C (10% wt.,100 mg) and the mixture was stirred under H₂ atmosphere for 3 hours. Theresulting mixture was filtered through a pad of celite and the filtratewas concentrated under vacuum to afford the title compound (400 mg).

MS_(—)2 (ESI): 230 [M+H]+

Intermediate 86 1,1-dimethylethyl{(1R)-1-methyl-2-[(2-{[3-(1-methylethyl)phenyl]oxy}-5-pyrimidinyl)amino]-2-oxoethyl}carbamate

To a solution of 2-{[3-(1-methylethyl)phenyl]oxy}-5-pyrimidinamine(Intermediate 85, 229 mg) in acetonitrile (20 mL) were addedN-{[(1,1-dimethylethyl)oxy]carbonyl}-D-alanine (378 mg, 2 mmol), HBTU(474 mg, 1.25 mmol) and DIPEA (387 mg, 3 mmol) and the mixture washeated at reflux and stirred overnight. The resulting mixture wasconcentrated under vacuum and water (100 mL) was added. It was extractedwith ethyl acetate (3 times 100 mL) and the combined organic layers weredried over sodium sulphate and evaporated to afford a brown oil, whichwas purified by silica gel chromatography (mobile phase: ethyl acetate:petroleum ether=1/5−1/2) to afford the title compound (300 mg).

Intermediate 87

N¹-(2-{[3-(1-methylethyl)phenyl]oxy}-5-pyrimidinyl)-D-alaninamide

To a solution of 1,1-dimethylethyl{(1R)-1-methyl-2-[(2-{[3-(1-methylethyl)phenyl]oxy}-5-pyrimidinyl)amino]-2-oxoethyl}carbamate(Intermediate 86, 300 mg) in ethyl acetate (50 mL) was bubbled HCl(gas). The mixture was stirred at room temperature for 1 hour. Theresulting mixture was concentrated under vacuum and neutralized with anaqueous saturated solution of NaHCO₃ to pH=8 and extracted withdichloromethane (5 times 30 mL). The combined organic layers were driedover sodium sulphate and evaporated to afford the title compound (200mg).

MS_(—)2 (ESI): 301 [M+H]+

Intermediate 88 ethyl 2-methyl-5-nitrophenyl ether

A mixture of 2-methyl-5-nitrophenol (450 mg, 2.94 mmol), ethyl iodide(356 μl, 4.41 mmol) and potassium carbonate (609 mg, 4.41 mmol) in 15 mLof acetone was heated at reflux for 2 days. The reaction mixture wasfiltrated, concentrated under vacuum, and the residue was partitionedbetween ethyl acetate and water. The organic layer was dried over sodiumsulfate and concentrated under vacuum to provide the title compound (397mg) which was directly used in the next step.

¹H NMR (400 MHz, methanol-d₄): δ ppm 7.78 (1H, dd), 7.72 (1H, d), 7.40(1H, dd), 4.22-4.17 (2H, q), 2.35 (3H, s), 1.53 (3H, t); UPLC: 0.82 min,182 [M+H]+.

Intermediate 89 3-(ethyloxy)-4-methylaniline

Fe powder (609 mg, 10.90 mmol) was added to a solution of ethyl2-methyl-5-nitrophenyl ether (Intermediate 88, 395 mg) in a mixtureTHF/water (15 mL/5 mL) followed by ammonium chloride (583 mg, 10.90mmol). The reaction mixture was stirred overnight under nitrogen. Thereaction mixture was poured into water (20 mL) and the iron wasfiltrated. Ethyl acetate was used to wash the solid filtrated. Thefiltrate was extracted with ethyl acetate (3 times). The combined ethylacetate layers were dried over sodium sulphate and concentrated to givethe title compound (303 mg) which was directly used in the next step.

¹H NMR (400 MHz, CDCl₃): δ ppm 6.90 (1H, d), 6.35-6.02 (2H, m), 3.97(2H, q), 2.11 (3H, s), 1.41 (3H, t); UPLC: 0.44 min, 152 [M+H]+.

Intermediate 90 3-(ethyloxy)-4-methylphenol

A suspension of 3-(ethyloxy)-4-methylaniline (Intermediate 89, 300 mg)in a mixture of water/concentrated sulfuric acid 98% (20 mL/7 mL) underargon was cooled at 0° C. A solution of sodium nitrite (151 mg, 2.182mmol) in 4 mL of water was slowly added. The reaction mixture wasstirred at 0° C. for 1 hour 30. The reaction mixture was then slowlyadded to a solution of water/concentrated sulfuric acid (18 mL/5 mL)pre-heated at 90° C. The reaction mixture was stirred at 90° C. for 1hour 15. After cooling down, the reaction mixture was extracted withethyl ether (4 times). The gathered organic phases were dried oversodium sulphate, filtered and concentrated under vacuum to afford thecompound (276 mg).

¹H NMR (400 MHz, methanol-d₄): δ ppm 6.87 (1H, d), 6.34 (1H, d), 6.25(1H, dd), 3.97 (2H, q), 2.06 (3H, s), 1.40-1.37 (3H, t); UPLC: 0.66 min,153 [M+H]+.

Intermediate 91 2-{[3-(ethyloxy)-4-methylphenyl]oxy}-5-nitropyrimidine

2-chloro-5-nitropyridine (114 mg, 0.716 mmol, 1 equiv) was dissolved in3 mL of dimethylformamide. 3-(ethyloxy)-4-methylphenol (Intermediate 90,109 mg) and potassium carbonate (198 mg, 1.432 mmol) were added. Thereaction mixture was stirred at room temperature 3 hours. The reactionmixture was filtered. The filtrated solid was washed withdichloromethane. The volatiles were evaporated under vacuum. Ethylacetate and brine were added to the residue. The compound was extracted2 times with ethyl acetate and 2 times with dichloromethane. Thegathered organic phases were dried over sodium sulphate, filtered andevaporated to afford the title compound which was used directly in thenext step (113 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 9.33 (2H, s), 7.21 (1H, d), 6.72-6.72(2H, m), 4.02 (2H, q), 2.25 (3H, s), 1.44 (3H, t); UPLC: 0.79 min, 276[M+H]+

Intermediate 92 2-{[3-(ethyloxy)-4-methylphenyl]oxy}-5-pyrimidinamine

Fe powder (89 mg, 1.598 mmol) was added to a solution of2-{[3-(ethyloxy)-4-methylphenyl]oxy}-5-nitropyrimidine (Intermediate 91,110 mg) in a mixture THF/water (9 mL/3 mL) followed by ammonium chloride(86 mg, 1.598 mmol). The reaction mixture was stirred overnight undernitrogen. The reaction mixture was poured into water and the iron wasfiltrated. Ethyl acetate was used to wash the solid filtrated. Thefiltrate was extracted with ethyl acetate (3 times). The combined ethylacetate layers were dried over sodium sulphate and concentrated to givethe title compound (94 mg) which was directly used in the next step.

UPLC: 0.64 min, 246 [M+H]+

Intermediate 931,1-dimethylethyl((1R)-1-{[(2-{[3-(ethyloxy)-4-methylphenyl]oxy}-5-pyrimidinyl)amino]carbonyl}propyl)carbamate

To a solution of(2R)-2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)butanoic acid (90 mg,0.445 mmol) in N,N-dimethylformamide (7 mL) DIPEA (97 μl, 0.557 mmol)and HATU (169 mg, 0.445 mmol) were added. The mixture reaction wasstirred during 10 minutes at room temperature, then2-{[3-(ethyloxy)-4-methylphenyl]oxy}-5-pyrimidinamine (Intermediate 92,91 mg) was added. The reaction mixture was stirred for 48 hours at roomtemperature. The reaction mixture was evaporated and purified by silicagel chromatography (Companion system) with a gradient Cyclohexane/EtOAcfrom 100/0 to 40/60 in 20 minutes and then 40/60 during 15 min to affordthe title compound (83 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 8.73 (2H, s), 7.13 (1H, d), 6.75-6.52(2H, m), 4.17-4.09 (1H, m), 3.98 (2H, q), 2.20 (3H, s), 2.04-1.88 (1H,m), 1.81-1.61 (1H, m), 1.46 (9H, s), 1.40 (3H, t), 1.02 (3H, t); UPLC:0.79 min, 431 [M+H]+

Intermediate 94(2R)-2-amino-N-(2-{[3-(ethyloxy)-4-methylphenyl]oxy}-5-pyrimidinyl)butanamide

To a solution of 1,1-dimethylethyl((1R)-1-{[(2-{[3-(ethyloxy)-4-methylphenyl]oxy}-5-pyrimidinyl)amino]carbonyl}propyl)carbamate(Intermediate 93, 80 mg) in dichloromethane (1 mL) cooled to 0° C., TFA(573 μl, 7.43 mmol) was added dropwise. The mixture reaction was stirredat 0° C. for 1.5. The solvent and the TFA were evaporated. The mixturewas diluted with dichloromethane and an aqueous saturated solution ofNaHCO3. The organic layer was separated, dried over sodium sulphate,filtered end evaporated to afford the title compound (65 mg) which wasdirectly used in the next step.

¹H NMR (400 MHz, CDCl₃): δ ppm 9.39 (1H, s), 8.82 (2H, s), 7.14 (1H, d),6.78-6.49 (2H, m), 3.98 (2H, q), 3.47 (1H, dd), 2.20 (3H, s), 2.09-1.91(1H, m), 1.81 (2H, sb), 1.76-1.56 (1H, m), 1.02 (3H, t); UPLC: 0.54 min,331 [M+H]+

Intermediate 95 N-{[(1,1-dimethylethyl)oxy]carbonyl}-3-methyl-D-valine

To a solution of 3-methyl-D-valine (900 mg, 6.86 mmol) in 7 mL of 1 Maqueous sodium hydroxide and 7 mL of methanol was added Boc-anhydride(1.797 g, 8.23 mmol) at 0° C. The reaction mixture was warmed to roomtemperature and stirred overnight. After most of the methanol wasevaporated, the solution was acidified to pH 2 with an aqueous solutionof HCl (1M) and extracted 3 times with ethylacetate (3×20 mL). Theorganic layers were combined and washed with brine (2×5 mL). Evaporationof the solvent afforded the title compound as a white solid with a 83%yield (1.36 g).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 12.44 (1H, s), 6.82 (1H, d), 3.76 (1H,d), 1.38 (9H, s), 0.93 (9H, s); UPLC: 0.64 min, 232 [M+H]+

Intermediate 961,1-dimethylethyl((1R)-2,2-dimethyl-1-{[(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)amino]carbonyl}propyl)carbamate

To a solution of N-{[(1,1-dimethylethyl)oxy]carbonyl}-3-methyl-D-valine(Intermediate 95, 20.1 mg) in dry N,Ndimethylformamide (1 mL), DIPEA(0.015 mL, 0.087 mmol) and then HATU (38.0 mg, 0.100 mmol) were addedand the reaction mixture was stirred for 15 minutes at room temperatureunder argon. Then 6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinamine(Intermediate 50, 10 mg) was added and the reaction mixture was stirredat 50° C. under argon 3 hours. The reaction was left at room temperatureovernight. The reaction mixture was evaporated. The residue obtained wasdissolved in dichloromethane. The organic phase was washed with brineand then with a saturated aqueous solution of NaHCO3. It was then driedover sodium sulphate. The residue obtained was purified on silica gel(Companion instrument) with cyclohexane/ethylacetate as eluents from100/0 to 70/30. This afforded the title compound (9.2 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 8.53 (1H, s), 8.21 (1H, d), 7.87 (1H, d),7.09 (1H, d),

6.76 (1H, d), 6.59 (2H, m), 5.43 (1H, d), 4.10 (1H, d), 3.75 (3H, s),2.17 (3H, s), 1.42 (9H, s), 1.07 (9H, s); UPLC_ipqc: 0.87 min, 444[M+H]+

Intermediate 973-methyl-N¹-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)-D-valinamide

To a solution of 1,1-dimethylethyl((1R)-2,2-dimethyl-1-{[(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)amino]carbonyl}propyl)carbamate(Intermediate 96, 8.2 mg) in dry dichloromethane (0.5 mL) cooled to 0°C., TFA (57 μl, 0.740 mmol) was added dropwise and the solution wasstirred for 3 hours at that temperature. The volatiles were evaporated.The residue was dissolved with dichloromethane (2 mL) and an aqueoussaturated solution of NaHCO3 was added (4 mL). The layers were separatedand the aqueous layer was extracted twice with dichloromethane. Thegathered organic layers were dried over sodium sulphate and evaporatedto afford the title compound (6.2 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 9.13 (1H, br s), 8.18 (2H, m), 7.11 (1H,d), 6.87 (1H, d), 6.60 (2H, m), 3.78 (3H, s), 3.27 (1H, s), 2.19 (3H,s), 1.69 (2H, br s), 1.06 (9H, s); UPLC_ipqc: 0.73 min, 344 [M+H]+

Intermediate 981,1-dimethylethyl((1R)-1-methyl-1-{[(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)amino]carbonyl}propyl)carbamate

To a solution of N-{[(1,1-dimethylethyl)oxy]carbonyl}-D-isovaline (94mg, 0.434 mmol) in dry N,N-Dimethylformamide (1 mL) DIPEA (0.114 mL,0.651 mmol) and HATU (165 mg, 0.434 mmol) were added. The reaction wasstirred at room temperature for 15 minutes.6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinamine (Intermediate 50,50 mg) was then added. After 1 hour of stirring at room temperature themixture was heated at 50° C. and stirred at that temperature for 4hours, it was then cooled down to room temperature and stirred overnightat that temperature. The mixture was quenched with brine (2 mL) andextracted with ethyl acetate (3×2 mL). Combined organic layers weredried over sodium sulphate, filtered and evaporated. The residue waspurified by flash chromatography on silica gel using a 10 g SNAP columnand cyclohexane/ethyl acetate as eluents from 100/0 to 60/40 (Biotagesystem) to afford the title compound as a white solid (65 mg).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 9.60 (1H, br s), 9.10 (1H, br s), 8.31(1H, br. s.), 8.03 (1H, br s), 7.12 (1H, d), 6.93 (1H, d), 6.69 (1H, d),6.53 (1H, dd), 3.74 (3H, s), 2.11 (3H, s), 1.72-1.86 (1H, m), 1.60-1.72(1H, m), 1.41 (9H, s), 1.33 (3H, s), 0.78 (3H, t); UPLC: 0.87 min, 430[M+H]+

Intermediate 99N¹-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)-D-isovalinamide

To a solution of 1,1-dimethylethyl((1R)-1-methyl-1-{[(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)amino]carbonyl}propyl)carbamate(Intermediate 98, 65 mg) in dry dichloromethane (3 mL) cooled to 0° C.,TFA (0.700 mL, 9.08 mmol) was added dropwise. The reaction was stirredat that temperature for 2 hours. The reaction was quenched with asaturated aqueous solution of NaHCO3 (20 mL) added at 0° C., andextracted with dichloromethane (3×7 mL). The combined organic layerswere dried over sodium sulphate, filtered and evaporated to afford thetitle compound as a white solid (44 mg).

Intermediate 1001-({[(1,1-dimethylethyl)oxy]carbonyl}amino)cyclobutanecarboxylic acid

To a solution of 1-aminocyclobutanecarboxylic acid (626 mg, 5.44 mmol)in 5.6 mL of 1 M aqueous sodium hydroxide and 4 mL of methanol was addedBoc-anhydride (1.425 g, 6.53 mmol) at 0° C. The reaction mixture waswarmed to room temperature and stirred for 12 hours. After most of themethanol was evaporated, the solution was acidified to pH 2 with 1 M HCland extracted with ethyl acetate. The organic extracts were combined andwashed with brine. Evaporation of the solvent afforded the titlecompound (1.09 g).

¹H NMR (400 MHz, DMSO-d6): δ ppm 12.21 (1H, s), 7.44 (1H, s), 2.29-2.47(2H, m), 2.09 (2H, q), 1.74-1.94 (2H, m), 1.36 (9H, s); UPLC: 0.56 min,216 [M+H]+

Intermediate 1011,1-dimethylethyl(1-{[(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)amino]carbonyl}cyclobutyl)carbamate

To a solution of1-({[(1,1-dimethylethyl)oxy]carbonyl}amino)cyclobutanecarboxylic acid(Intermediate 100, 70.1 mg) in dry N,N-dimethylformamide (2 mL) DIPEA(0.095 mL, 0.543 mmol) and TBTU (112 mg, 0.347 mmol) were added. Thereaction mixture was stirred at room temperature for 15 minutes,6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinamine (Intermediate 50,50 mg) was then added and the mixture was stirred at room temperaturefor 1 day. The reaction was quenched with water (5 mL) and extractedwith ethyl acetate (3×5 mL). The combined organic layer was washed withbrine (3×8 mL), separated, dried over sodium sulphate, filtered andevaporated. The residue was purified by flash chromatography on silicagel using a 25 g SNAP column and cyclohexane:ethylacetate as eluentsfrom 10:0 to 7:3. This afforded the title compound as a white powder (80mg).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 9.52 (1H, s), 8.34 (1H, s), 7.96 (1H,m), 7.51 (1H, s), 7.13 (1H, d), 6.95 (1H, d), 6.70 (1H, d), 6.54 (1H,dd), 3.75 (3H, s), 2.13 (3H, s), 2.11 (2H, br. s.), 1.76-1.97 (2H, m),1.39 (9H, s), 1.26 (2H, s); UPLC_B: 0.93 min, 426 [M−H]−

Intermediate 1021-amino-N-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)cyclobutanecarboxamide

To a solution of 1,1-dimethylethyl(1-{[(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)amino]carbonyl}cyclobutyl)carbamate(Intermediate 101, 80 mg) in dry dichloromethane (2 mL) cooled to 0° C.TFA (0.865 mL, 11.23 mmol) was added dropwise. The reaction mixture wasstirred at 0° C. for 2 hours, it was then allowed to reach roomtemperature. Dichloromethane and the excess of TFA were evaporated. Theresidue was diluted with dichloromethane (5 mL) and neutralized with asaturated solution of NaHCO3. The organic layer was separated, driedover sodium sulphate, filtered and evaporated to afford the tilte as awhite solid (60 mg).

¹H NMR (400 MHz, CDCl₃) d ppm 9.43 (1H, br. s.), 8.37-7.89 (2H, m), 7.09(1H, d), 6.85 (1H, d), 6.67-6.39 (2H, m), 3.77 (3H, s), 2.92-2.62 (2H,m), 2.18 (3H, s), 1.97-2.06 (2H, m), 1.97-1.74 (2H, m); UPLC: 0.54 min,328 [M+H]+

Intermediate 1031-({[(1,1-dimethylethyl)oxy]carbonyl}amino)cyclopropanecarboxylic acid

The title compound (998 mg) was made in a similar fashion to thepreparation of Intermediate 100 replacing 1-aminocyclobutanecarboxylicacid with 1-aminocyclopropanecarboxylic acid (550 mg, 5.44 mmol).

¹H NMR (400 MHz, DMSO-d6): δ ppm 12.26 (1H, s), 7.40 (1H, s), 1.38 (9H,s), 1.26 (2H, m), 0.96 (2H, m); UPLC: 0.52 min, 202 [M+H]+.

Intermediate 1041,1-dimethylethyl(1-{[(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)amino]carbonyl}cyclopropyl)carbamate

The title compound (80 mg) was made in a similar fashion to thepreparation of Intermediate 101 replacing1-({[(1,1-dimethylethyl)oxy]carbonyl}amino)cyclobutanecarboxylic acidwith 1-({[(1,1-dimethylethyl)oxy]carbonyl}amino)cyclopropanecarboxylicacid (Intermediate 103, 65.5 mg).

¹H NMR (400 MHz, DMSO-d₆) d ppm 9.56-9.76 (1H, m), 8.34 (1H, d), 8.04(1H, dd), 7.31-7.51 (1H, m), 7.13 (1H, d), 6.94 (1H, d), 6.71 (1H, d),6.55 (1H, dd), 3.75 (3H, s), 2.13 (3H, s), 1.42 (9H, s), 1.26-1.39 (2H,m), 0.75-1.07 (2H, m); UPLC_B: 0.89 min, 412 [M−H]−

Intermediate 1051-amino-N-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)cyclopropanecarboxamide

The title compound (58 mg) was made in a similar fashion to thepreparation of Intermediate 102 replacing 1,1-dimethylethyl(1-{[(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)amino]carbonyl}cyclobutyl)carbamatewith 1,1-dimethylethyl(1-{[(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)amino]carbonyl}cyclopropyl)carbamate(Intermediate 104, 80 mg)

¹H NMR (400 MHz, CDCl₃) d ppm 9.85 (1H, s), 8.23 (1H, d), 8.14 (1H, dd),7.09 (1H, d), 6.85 (1H, d), 6.67-6.45 (2H, m), 3.77 (3H, s), 2.18 (3H,s), 1.85 (2H, br. s.), 1.47-1.57 (2H, m), 0.96-0.85 (2H, m); UPLC: 0.52min, 314 [M+H]+

Intermediate 1063-(6-fluoro-3-pyridinyl)-5,5-dimethyl-2,4-imidazolidinedione

Two reactions were set up in parallel. For each of them, to a solutionof 5,5-dimethyl-2,4-imidazolidinedione (1.5 g, 11.71 mmol) in drydichloromethane (100 mL), (6-fluoro-3-pyridinyl)boronic acid (1.980 g,14.05 mmol), copper(II) acetate (2.126 g, 11.71 mmol) and pyridine(1.420 mL, 17.56 mmol) were added. The reaction was left under stirringunder air atmosphere at room temperature overnight. The two reactionmixtures were combined and the solid was filtered off. The resultingsolution was washed with water (90 mL). The aqueous phase was extractedtwice with dichloromethane (twice 90 mL). The organic phases werecombined and washed with brine and dried over anhydrous sodium sulphate.Removal of the solvent afforded a residue which was purified by silicagel chromatography (Biotage system, 100 g SNAP column) using as eluent agradient and cyclohexane/ethyl acetate from 65/35 to 50/50 to afford thetitle compound as a white solid (1.27 g).

¹H NMR (400 MHz, CDCl₃): δ ppm 8.40 (1H, m), 7.94 (1H, m), 7.06 (1H,dd), 6.18 (1H, br. s), 1.57 (6H, s); UPLC_ipqc: 0.56 min, 224 [M+H]+

Intermediate 107 2-methylalaninate hydrochloride

2-amine-2-methylpropionic acid (25 g, 242.43 mmol) was dissolved inmethanol (150 mL). Thionyl chloride (25 mL) was added dropwise at 0° C.to the reaction mixture. The reaction was refluxed for 3 hours,evaporated and dried under vacuum. The solid was washed several timeswith Et₂O and dried, to afford the title compound (37 g) as a whitesolid.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.71 (3H, s), 3.77 (3H, s), 1.5 (6H,s).

Intermediate 108 3-(1,1-dimethylethyl)-4-hydroxybenzaldehyde

2-(1,1-dimethylethyl)phenol (10 g, 66.67 mmol) was dissolved in 40 mL ofMeOH and NaOH (40 g, 1 mol) dissolved in 40 mL of water was addeddropwise. Then 40 mL of CHCl₃ was added (during the course of 1 h) at60° C. The reaction mixture was stirred at that temperature for 3 h.After cooling down to r.t., the mixture was cooled to 0° C. and 4M HClwas added until the solution reached pH 5-6. The mixture was extractedwith DCM (three times) and the collected organic were dried over Na₂SO₄,filtered and evaporated. The crude was charged on a silica gel columnand eluted with Cyclohexane/EtOAc (from 100:0 to 80:20Cyclohexane/EtOAc, then plateau at 80:20) affording 766 mg of the of thetitle compound.

¹H-NMR (400 MHz, DMSO-d₆): δ ppm 10.62 (1H, s), 9.79 (1H, s), 7.73 (1H,br. s), 7.67-7.57 (1H, m), 7.01-6.90 (1H, m), 1.38 (9H, s); UPLC_ipqc:0.97 min, 177 [M−H]⁻.

Intermediate 109 3-(1,1-dimethylethyl)-4-hydroxybenzonitrile

3-(1,1-dimethylethyl)-4-hydroxybenzaldehyde (Intermediate 108, 550 mg)and hydroxylamine hydrochloride (322 mg, 4.63 mmol) were stirred in 8.0mL of acetic acid at reflux for 1 h. After cooling down to 0° C., themixture was poured into Et₂O and washed once with water and once withNaOH (5% aqueous solution). The collected aqueous phases were extractedwith Et₂O (two times) and the combined organic phases were dried overNa₂SO₄, filtered, evaporated and triturated with pentane affording 540mg of the title compound.

¹H-NMR (400 MHz, DMSO-d₆): δ ppm 10.92 (1H, br. s), 7.53-7.45 (2H, m),6.92 (1H, d), 1.34 (9H, s); UPLC_ipqc: 1.03 min, 174 [M−H]⁻.

Intermediate 110 3-bromo-4-[(phenylmethyl)oxy]benzonitrile

3-bromo-4-hydroxybenzonitrile (5.94 g, 0.03 mol) was dissolved in 100 mLof dry acetone. Potassium carbonate (8.29 g, 0.06 mol) was added. To thereaction mixture, benzyl bromide was then added dropwise (5 g, 0.03mol). The reaction mixture was stirred at 50° C. overnight. Then it wascooled down to room temperature, filtered and evaporated. The residueobtained was dissolved in ethyl acetate (300 mL) and water was added(200 mL). The phases were separated and the gathered organic phases weredried over sodium sulphate. Evaporation led to the title compound (7.6g) as a light yellow solid.

¹H NMR (400 MHz, CDCl₃): δ ppm 7.85 (1H, d), 7.57 (1H, dd), 7.45-7.36(5H, m), 6.99 (1H, d), 5.23 (2H, s); UPLC_ipqc: 1.24 min.

The following compounds were prepared using the foregoing methodology,replacing 3-bromo-4-hydroxybenzonitrile with the appropriatelysubstituted phenol, as described in the foregoing Reaction Schemes.

UPLC_ipqc nt. Structure Name Phenol NMR characterizationcharacterization 11

1-bromo-4- [(phenylmethyl) oxy]-2- [(trifluoromethyl) oxy]benzene4-bromo-3- [(trifluoromethyl) oxy]phenol ¹H NMR (400 MHz, CDCl₃): δ ppm7.19-7.78 (6H, m), 6.73-7.14 (2H, m), 5.06 (2H, br. s.) 1.44 min

Intermediate 112 3-ethyl-4-[(phenylmethyl)oxy]benzonitrile

To a 1M solution of ethyl magnesium bromide (2 mL, 1.5 equiv), THF (10mL) was added and the reaction mixture was cooled down to 0° C. Asolution of 0.5 M in THF of zinc dichloride (4 mL, 1.5 equiv) was addedslowly and the reaction mixture was stirred for 30 min at the sametemperature. Pd(tBu₃P)₂ (102 mg, 0.1 equiv) was added followed by theaddition of a solution of 3-bromo-4-[(phenylmethyl)oxy]benzonitrile(Intermediate 110, 576 mg) in THF and the reaction was allowed to reachroom temperature. After stirring 30 minutes, some additional Pd(tBu₃P)₂was added (51 mg, 0.05 equiv), then 30 min stirring, then third additionof Pd(tBu₃P)₂ (51 mg, 0.05 equiv) and stirring 30 min. The reactionmixture was quenched with an aqueous saturated solution of NH₄Cl (100mL) and extracted 3 times with ethyl acetate (3×150 mL). The gatheredorganic phases were filtrated on celite, dried over sodium sulphate. Theresidue obtained was purified by chromatography on silica gel (Companionsystem, 40 g Si cartridge) using as eluent a gradient cyclohexane/ethylacetate 100:0, then 100:0 to 90:10. Evaporation afforded the titlecompound (336 mg) as a white solid.

¹H NMR (400 MHz, CDCl₃): δ ppm 7.49-7.27 (7H, m), 6.94 (1H, d), 5.15(2H, s), 2.72 (2H, q), 1.23 (3H, t); UPLC_ipqc: 1.30 min, 236 [M−H]−.

Intermediate 1131-methyl-4-[(phenylmethyl)oxy]-2-[(trifluoromethyl)oxy]benzene

Preparation of organometallic solution: To a solution of 1M ZnCl₂ inEt₂O (6 mL) was slowly added at room temperature a solution of 1.4Mmethyl magnesium bromide solution in THF (4.3 mL) and the reactionmixture was stirred for 20 minutes at room temperature.

To a solution of1-bromo-4-[(phenylmethyl)oxy]-2-[(trifluoromethyl)oxy]benzene(Intermediate 111, 537 mg, 1.55 mmol) and Pd(tBu₃P)₂ (208 mg, 0.4 mmol),warmed at 60° C., were added 5.15 mL of the organometallic solutionpreviously formed and the reaction mixture was stirred for 1 hour at 60°C. Further 5.15 mL of the organometallic solution were added and thereaction mixture was stirred for 30 minutes at 60° C. After cooling thereaction was quenched with water (1 mL), diluted with an aqueoussaturated solution of ammonium chloride (20 mL) and extracted with ethylacetate (2×50 mL). The organic layer was washed with brine (2×20 mL),dried (Na₂SO₄), filtered and evaporated. The crude was purified by flashchromatography (silica, from 100:0 to 80:20 Cyclohexane/EtOAc) to givethe title compound (203 mg) as solid.

¹H NMR (400 MHz, CDCl₃): δ ppm 7.30-7.49 (5H, m), 7.15 (1H, d),6.79-6.90 (2H, m), 5.05 (2H, s), 2.25 (3H, s); UPLC_ipqc: 1.43 min, 281[M−H]−.

The following compounds were prepared using the foregoing methodology,replacing 1-bromo-4-[(phenylmethyl)oxy]-2-[(trifluoromethyl)oxy]benzenewith the appropriately substituted halo compound, as described in theforegoing Reaction Schemes.

Halo UPLC_ipqc nt. Structure Name compound NMR characterizationcharacterization 14

3-methyl-4-[(5- nitro-2- pyrimidinyl)oxy] benzonitrile 3-bromo-4-[(5-nitro-2- pyrimidinyl)oxy] benzonitrile] (Intermediate 130) ¹H NMR (400MHz, CDCl₃): δ ppm 9.36 (2H, s), 7.68 (1H, s), 7.64 (1H, d), 7.25 (1H,d), 2.26 (3H, s) 0.94 min

Intermediate 115 3-ethyl-4-hydroxybenzonitrile

3-ethyl-4-[(phenylmethyl)oxy]benzonitrile (Intermediate 112, 334 mg) wasdissolved in 15 mL of EtOAc/EtOH (2/1) and 10% mol Pd/C (0.1 equiv) wasadded to the solution. The resulting mixture was stirred overnight atroom temperature under hydrogen gas atmosphere. The reaction mixture wasfiltered under argon and the solvent was removed. The residue obtainedwas purified by chromatography on silica gel (Companion system, 40 g Sicartridge) using as eluent a gradient cyclohexane/ethyl acetate from100:0 to 80:20. Evaporation afforded the title compound (148 mg) as awhite solid.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 10.50 (1H, br s), 7.50 (2H, m), 6.92(1H, d), 2.55 (2H, q), 1.12 (3H, t); UPLC_ipqc: 0.84 min, 146 [M−H]−

The following compounds were prepared using the foregoing methodology,replacing 3-ethyl-4-[(phenylmethyl)oxy]benzonitrile with the appropriatebenzylated phenol.

Benzylated UPLC_ipqc nt. Structure Name phenol NMR characterizationcharacterization 16

4-methyl-3- [(trifluoromethyl) oxy]phenol 1-methyl-4- [(phenylmethyl)oxy]-2- [(trifluoromethyl) oxy] (Intermediate 113) ¹H NMR (400 MHz,DMSO- d₆): δ ppm 9.77 (1H, br. s.), 7.15 (1H, d), 6.63-6.75 (2H, m),2.13 (3H, s) 1.01 min, 192 [M − H]−

Intermediate 117 4-hydroxy-2-iodobenzonitrile

To a solution of 2-fluoro-4-iodobenzonitrile (5.0 g, 20.24 mmol) in dryacetonitrile (100 mL) potassium trimethylsilanolate (1.18 g) was addedand the reaction mixture was stirred overnight at 50° C. The solvent wasremoved under reduced pressure and the residue was dissolved in ethylacetate (100 mL) and an aqueous pH 3 buffer solution was added up to pH˜5. Two phases were separated and the organic layer was dried (Na₂SO₄),filtered and evaporated to afford the title compound (4.90 g) as brownsolid.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 10.92 (1H, s), 7.65 (1H, d), 7.39 (1H,d), 6.93 (1H, dd); UPLC_ipqc: 0.81 min, 244 [M−H]−.

The following compounds were prepared using the foregoing methodology,replacing 4-fluoro-2-iodobenzonitrile with the appropriately substitutedfluoro benzonitrile, as described in the foregoing Reaction Schemes.

UPLC_ipqc Fluoro character- nt. Structure Name benzonitrile ization 18

4-hydroxy-3- methyl- benzonitrile 4-fluoro-3- methyl- benzonitrile 0.74min, 134 [M + H]+, 132 [M − H]−

Intermediate 119 4-hydroxy-2-[(trifluoromethyl)oxy]benzonitrile

Two reactions were carried out in parallel (A and B) and then the tworeaction mixtures were combined to run work-up and purification.

Reaction A: To a solution of 4-Methoxy-2-(trifluoromethoxy)benzonitrile(50 mg, 0.23 mmol) in 1,2-dichloroethane (1 mL) was added 1M BBr₃solution in DCM (0.69 mL, 0.69 mmol) dropwise. The resulting reactionmixture was stirred under microwave irradiation five times (setparameters: T=100° C., t=1 hour) adding further 1M BBr₃ solution in DCM(1 mL) each time. The total amount of 1M BBr₃ solution in DCM used was4.69 mL.

Reaction B: In a vial were added4-Methoxy-2-(trifluoromethoxy)benzonitrile (750 mg, 3.45 mmol),1,2-dichloroethane (5 mL) and then 1M BBr₃ solution in DCM (10.36 mL,10.36 mmol) dropwise. The resulting reaction mixture was stirred undermicrowave irradiation for 1 hour (set T=100° C.). To the reactionmixture further 1M BBr₃ solution in DCM (1 mL) was added and theresulting reaction mixture was stirred under microwave irradiation threemore times (set parameters: T=100° C., t=1.5 hours), adding further 1MBBr₃ solution in DCM (0.8 mL) each time. The total amount of 1M BBr₃solution in DCM used was 13.76 mL.

The two reactions mixtures A and B were added dropwise to a NaHCO₃saturated aqueous solution and the pH was adjusted to 7 with theaddition of solid NaHCO₃. The two phases were separated and the aqueousphase was extracted with DCM (1×) and with EtOAc (2×). The combinedorganic phases were dried and evaporated to dryness to give the titlecompound in mixture with unreacted starting material (1.48 g) as a blackoil. This mixture was used in the next step without furtherpurification.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 11.35 (1H, s), 7.82 (1H, d), 6.91-6.98(2H, m); UPLC_ipqc: 0.88 min, 204 [M+H]+, 202 [M−H]−.

Intermediate 1204-[(5-nitro-2-pyridinyl)oxy]-3-(trifluoromethyl)benzonitrile

A mixture of 2-chloro-5-nitropyridine (70 mg, 0.44 mmol),4-hydroxy-3-(trifluoromethyl)benzonitrile (91 mg, 0.49 mmol), K₂CO₃ (92mg, 0.66 mmol) in DMF (2 mL) was stirred at 50° C. overnight. Water (4mL) was added and a precipitate was formed. The solid was filtered-offand it was triturated with MeOH to give the title compound (85 mg) as abrownish solid.

¹H NMR (400 MHz, CDCl₃): δ ppm 8.99 (1H, d), 8.60 (1H, dd), 8.07 (1H,s), 7.95 (1H, d), 7.48 (1H, d), 7.19-7.32 (1H, m); UPLC_ipqc: 1.1 min,310 [M+H]+.

The following compounds were prepared using the foregoing methodology,reacting the appropriate halo nitroaryl such as2-chloro-5-nitropyridine, 2-chloro-5-nitropyrimidine,1-fluoro-4-nitrobenzene etc. with the appropriately substituted phenolat a suitable temperature, optionally under microwave irradiation, asdescribed in the foregoing Reaction Schemes. Some final products werepurified by flash-chromatography (Silica; Cyclohexane/EtOAc or otherappropriate solvent system).

Halo NMR UPLC_ipqc Int. Structure Name nitroaryl Phenol characterizationcharacterization 121

2-iodo-4- [(5-nitro-2- pyrimidinyl) oxy]benzonitrile 2-chloro- 5-nitropyrimidine 4-hydroxy- 2- iodobenzonitrile (Intermediate 117) ¹H-NMR(400 MHz, DMSO- d₆): δ ppm 9.50 (2H, br. s), 8.14 (1H, br. s), 8.05-7.99 (1H, m), 7.57-7.63 (1H, m) 1.03 min, 369 [M + H]⁺ 122

3-bromo-4- [(5-nitro-2- pyridinyl) oxy]benzonitrile 2-chloro- 5-nitropyridine 3-bromo-4- hydroxy- benzonitrile ¹H-NMR (400 MHz, CDCl₃):δ ppm 9.01 (1 H, br. s), 8.65- 8.56 (1 H, m), 8.02 (1H, s), 7.75 (1 H,d), 7.38 (1H, d), 7.25 (1H, d) 1.08 min, 320 [M]+, Br pattern 123

3-(1,1- dimethylethyl)- 4-[(5- nitro-2- pyridinyl) oxy]benzonitrile2-chloro- 5- nitropyridine 3-(1,1- dimethylethyl)- 4-hydroxybenzonitrile (Intermediate 109) ¹H NMR (400 MHz, DMSO- d₆): δ ppm9.09-9.04 (1H, m), 8.74- 8.64 (1H, m), 7.87 (1H, br. s), 7.83- 7.77 (1H,m), 7.44 (1H, d), 7.33 (1H, d), 1.32 (9H, s) 1.23 min, 298 [M + H]⁺ 124

2-({4- methyl-3- [(trifluoromethyl) oxy]phenyl} oxy)-5- nitropyridine2-chloro- 5- nitropyridine 4-methyl-3- [(trifluoromethyl) oxy]phenol ¹HNMR (400 MHz, CDCl₃): δ ppm 9.02 (1 H, d), 8.49 (1 H, dd), 7.33 (1 H,d), 6.97- 7.14 (3 H, m), 2.34 (3 H, s) 1.29 min, 315 [M + H]⁺. 125

2- (methyloxy)- 4-[(5- nitro-2- pyridinyl) oxy]benzonitrile 2-chloro- 5-nitropyridine 4-hydroxy- 2- (methyloxy) benzonitrile ¹H NMR (400 MHz,CDCl₃): δ ppm 9.05 (1H, dd) 8.56 (1H, dd) 7.65 (1H, d) 7.14 (1H, dd)6.80- 6.87 (2H, m) 3.94 (3H, s) 0.99 min, 272 [M + H]+ 126

2- (methyloxy)- 4-[(4- nitrophenyl) oxy]benzonitrile 1-fluoro- 4-nitrobenzene 4-hydroxy- 2- (methyloxy) benzonitrile ¹H NMR (400 MHz,CDCl₃): δ ppm 8.24- 8.35 (2H, m) 7.59 (1H, d) 7.10-7.20 (2H, m) 6.69(1H, d) 6.65 (1H, dd) 3.92 (3H, s) 1.10 min 127

2-hydroxy- 4-[(4- nitrophenyl) oxy]benzonitrile 1-fluoro- 4-nitrobenzene 2,4- dihydroxybenzonitrile ¹H NMR (400 MHz, DMSO- d₆): δppm 11.41 (1H, br. s.) 8.26-8.34 (2H, m) 7.71 (1H, d) 7.25- 7.40 (2H, m)6.69 (1H, dd) 6.67 (1H, d) 0.97 min, 255 [M − H]− 128

2-hydroxy- 4-[(5-nitro- 2- pyridinyl) oxy]benzonitrile 2-chloro- 5-nitropyridine 2,4- dihydroxybenzonitrile ¹H NMR (400 MHz, DMSO- d₆): δppm 11.45(1H, br. s.), 9.06 (1H, d), 8.66 (1H, dd), 7.72 (1H, d), 7.34(1H, d), 6.77-6.85 (2H, m) 0.87 min, 256 [M − H]− 129

2-iodo-4- [(5-nitro-2- pyridinyl) oxy]benzonitrile 2-chloro- 5-nitropyridine 4-hydroxy- 2- iodobenzonitrile (Intermediate 117) 1H NMR(400 MHz, DMSO- d6): δ ppm 9.08 (1H, d), 8.70 (1H, dd), 8.03 (1H, d),7.98 (1H, d), 7.52 (1H, dd), 7.41 (1H, d) 1.10 min 130

3-bromo-4- [(5-nitro-2- pyrimidinyl) oxy]benzonitrile 2-chloro- 5-nitropyrimidine 3-bromo-4- hydroxybenzonitrile ¹H NMR (400 MHz, DMSO-d₆): δ ppm 9.51 (2H, s), 8.44-8.47 (1H, m), 8.03- 8.10 (1H, m), 7.74(1H, d). 131

3-methyl- 4-[(5-nitro- 2- pyridinyl) oxy]benzonitrile 2-chloro- 5-nitropyridine 4-hydroxy- 3- methylbenzonitrile (Intermediate 118) ¹H NMR(400 MHz, CDCl₃): δ ppm 9.02 (1H, d), 8.57 (1H, dd), 7.50- 7.71 (2H, m),7.13- 7.25 (2H, m), 2.23 (3 H, s) 1.04 min, 256 [M + H]+. 132

4-[(5-nitro- 2- pyridinyl) oxy]-2- [(trifluoromethyl) oxy]benzonitrile2-chloro- 5- nitropyridine 4-hydroxy- 2- [(trifluoromethyl)oxy]benzonitrile (Intermediate 119) ¹H NMR (400 MHz, CDCl₃): δ ppm 9.02-9.11 (1H, m), 8.55-8.65 (1H, m), 7.82 (1 H, d), 7.25- 7.35 (2 H, m),7.20 (1 H, d) 1.14 min, 326 [M + H]+

Intermediate 133 2-ethyl-4-[(5-nitro-2-pyrimidinyl)oxy]benzonitrile

In a flamed 2-necked flask, under N₂, to a solution of ZnCl₂ (0.82 mL ofa 0.5 M solution on THF, 0.41 mmol) in 1.0 mL of THF, cooled at −15° C.,EtMgBr (0.41 mL of a 1.0 M solution in THF, 0.41 mmol) was slowly addedand the reaction mixture was stirred at that temperature for 1 h. ThenPd(tBu₃P)₂ (7.0 mg, 0.03 mmol) was added, followed by2-iodo-4-[(5-nitro-2-pyrimidinyl)oxy]benzonitrile (Intermediate 121,50.0 mg) in THF (1.0 mL) and the reaction mixture was stirred at −15° C.for 1 h and then the NaCl/ice bath was removed. After 2 hours at r.t.additional 3.5 mg (0.015 mmol) of (tBu₃P)₂ were added. The reaction wasquenched with NH₄Cl (saturated aqueous solution) and extracted withEtOAc (three times). The collected organic were dried over Na₂SO₄,filtered and evaporated. The residue obtained was charged on a silicagel column and eluted with Cyclohexane/EtOAc (from 100:0 to 90:10Cyclohexane/EtOAc, plateau at 90:10) affording 25 mg of the titlecompound.

¹H-NMR (400 MHz, DMSO-d₆): δ ppm 9.47 (2H, br. s), 7.94 (1H, d), 7.49(1H, br. s), 7.41-7.33 (1H, m), 2.84 (2H, q), 1.24 (3H, t); UPLC_ipqc:1.06 min, 271 [M+H]⁺.

Intermediate 134 2-cyclopropyl-4-[(5-nitro-2-pyridinyl)oxy]benzonitrile

Preparation of organometallic solution: to a solution of 0.5M ZnCl₂ inTHF (9 mL) a solution of 0.5M Cyclopropyl Magnesium bromide in THF (9mL) was slowly added at r.t. and the reaction mixture was stirred for 20minutes at r.t.

To a solution of 2-iodo-4-[(5-nitro-2-pyridinyl)oxy]benzonitrile(Intermediate 129, 550 mg) and Pd(tBu₃P)₂ (76 mg, 0.15 mmol), warmed at60° C., were added 6 mL of the organometallic solution previously formedand the reaction mixture was stirred for 1 hour at 60° C. Further 6 mLof the organometallic solution were added and the reaction mixture wasstirred for additional 1 hour at 60° C. Further 6 mL of theorganometallic solution were added and the reaction mixture was stirredfor additional 1 hour at 60° C. After cooling the reaction was quenchedwith water (1 mL), diluted with an aqueous saturated solution ofammonium chloride (20 mL) and extracted with ethyl acetate (2×50 mL).The organic layer was washed with brine (2×20 mL), dried (Na₂SO₄),filtered and evaporated and the residue was purified by flashchromatography on silica gel (SNAP 50 g), eluting from 100:0 to 80:20n-hexane/ethyl acetate affording the title compound (400 mg) as whitesolid.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 9.06 (1H, d), 8.67 (1H, dd), 7.88 (1H,d), 7.35 (1H, d), 7.23 (1H, dd), 7.01 (1H, dd), 2.17-2.27 (1H, m),1.10-1.19 (2H, m), 0.82-0.90 (2H, m); UPLC_ipqc: 1.13 min, 282 [M+H]+.

The following compounds were prepared using the foregoing methodology,replacing Cyclopropyl Magnesium bromide with the appropriate Grignardreagent to form the organozinc reagent, as described in the foregoingReaction Schemes.

Grignard UPLC_ipqc Int. Structure Name reagent NMR characterizationcharacterization 135

2-ethyl-4-[(5- nitro-2- pyridinyl)oxy] benzonitrile Ethyl magnesiumbromide ¹H-NMR (400 MHz, CDCl₃): δ ppm 9.06 (1H, d) 8.56 (1H, dd) 7.72(1H, d) 7.18 (1H, d) 7.10-7.17 (2H, m) 2.95 (2H, q) 1.35 (3H, t) 1.12min, 270 [M + H]+ 136

3-ethyl-4-[(5- nitro-2- pyrimidinyl)oxy] benzonitrile Ethyl Magnesiumbromide ¹H NMR (400 MHz, CDCl₃): δ ppm 9.35 (2H, s), 7.70 (1H, s), 7.64(1H, dd), 7.23 (1H, d), 2.62 (2H, q), 1.25 (3H, t) 1.02 min

Intermediate 137 3-cyclopropyl-4-[(5-nitro-2-pyridinyl)oxy]benzonitrile

In a vial 3-bromo-4-[(5-nitro-2-pyridinyl)oxy]benzonitrile (Intermediate122, 800 mg) was dissolved in 16.0 mL of toluene. Cyclopropylboronicacid (1073.8 mg, 12.5 mmol) was added, followed by Pd(OAc)₂ (56.1 mg,0.25 mmol) and (Cy)₃P (70.0 mg 0.25 mmol). Then, an aqueous solution(8.0 mL of water) of K₃PO₄ (1855.0 mg, 8.75 mmol) was added. Thereaction mixture was heated at 80° C. overnight. After cooling down tor.t., the mixture was partitioned between brine and EtOAc and theseparated aqueous phase was extracted with EtOAc (three times). Thecollected organic were dried over Na₂SO₄, filtered and evaporated. Thecrude obtained was charged on a silica gel column and eluted withCyclohexane/EtOAc (from 100:0 to 80:20 Cyclohexane/EtOAc) affording 634mg of the title compound.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 9.04 (1H, br. s), 8.69 (1H, dd), 7.75(1H, d), 7.58 (1H, s), 7.41 (2H, t), 1.90-1.80 (1H, m), 0.90-0.73 (4H,m); UPLC_ipqc: 1.12 min, 282 [M+H]⁺.

Intermediate 1382-(1-methylethenyl)-4-[(5-nitro-2-pyridinyl)oxy]benzonitrile

To a solution of 2-iodo-4-[(5-nitro-2-pyridinyl)oxy]benzonitrile(Intermediate 129, 5.0 g) in DMF (50 mL) were added K₃PO₄ (5.77 g, 27.24mmol), Pd(tBu₃)₂ (696 mg, 1.36 mmol) and4,4,5,5-tetramethyl-2-(1-methylethenyl)-1,3,2-dioxaborolane (3.84 mL,20.43 mmol) and the reaction mixture was stirred for 4 hours at 110° C.After cooling the reaction was diluted with water (100 mL) and extractedwith ethyl acetate (3×100 mL). The organic layer was washed with icecold brine (3×50 mL), dried (Na₂SO₄), filtered and evaporated and theresidue was purified by flash chromatography on silica gel (SNAP 100 g)eluting from 100:0 to 80:20 cyclohexane/ethyl acetate to afford thetitle compound (1.8 g) as white solid.

¹H-NMR (400 MHz, DMSO-d₆): δ ppm 9.08 (1H, d), 8.69 (1H, dd), 7.97 (1H,d), 7.47 (1H, d), 7.40 (2H, d), 5.46 (1H, s), 5.32 (1H, s), 2.16 (3H,s); UPLC_ipqc: 1.14 min, 282 [M+H]+.

Intermediate 139 2-(ethyloxy)-4-[(4-nitrophenyl)oxy]benzonitrile

2-hydroxy-4-[(4-nitrophenyl)oxy]benzonitrile (Intermediate 127, 87.2 mg)was dissolved in DMF (5 mL). K₂CO₃ (92.2 mg, 0.67 mmol) and iodoethane(32 μL, 0.40 mmol) were added. The reaction mixture was stirred at r.t.After 16 h, the reaction mixture was evaporated to dryness to give thecrude product that was purified by silica gel chromatography (from 100:0to 50:50

Cyclohexane/EtOAc in 10 CV; then 50:50 Cyclohexane/EtOAc for 10 CV) toobtain 84.9 mg of the title compound as a yellow solid.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.26-8.35 (2H, m) 7.82 (1H, d)7.26-7.35 (2H, m) 7.09 (1H, d) 6.80 (1H, dd) 4.18 (2H, q) 1.36 (3H, t);UPLC_ipqc: 1.16 min, [M+H]+.

The following compounds were prepared using the foregoing methodology,replacing 2-hydroxy-4-[(4-nitrophenyl)oxy]benzonitrile with theappropriately substituted phenol and iodoethane with the appropriateelectrophile, as described in the foregoing Reaction Schemes. Some finalproducts were purified by flash-chromatography (Silica;Cyclohexane/EtOAc or other appropriate solvent system).

UPLC_ipqc Electro- character- Int. Structure Name Phenol phile NMRcharacterization ization 140

2- [(cyclopropyl- methyl) oxy]-4- [(5-nitro-2- pyridinyl)oxy]benzonitrile 2-hydroxy-4- [(5-nitro-2- pyridinyl)oxy] benzonitrile(Intermediate 128) (bromo- methyl) cyclo- propane ¹H NMR (400 MHz,DMSO-d₆): δ ppm 9.06 (1H, d), 8.67 (1H, dd), 7.83 (1H, d), 7.36 (1H, d),7.19 (1H, d), 6.97 (1H, dd), 3.97 (2H, d), 1.17-1.35 (1H, m), 0.53- 0.65(2H, m), 0.29- 0.40 (2H, m) 1.14 min, 312 [M + H]+ 141

2-(ethyloxy)-4- [(5-nitro-2- pyridinyl)oxy] benzonitrile 2-hydroxy-4-[(5-nitro-2- pyridinyl)oxy] benzonitrile (Intermediate 128) iodoethane¹H NMR (400 MHz, DMSO-d₆): δ ppm 9.06 (1H, d), 8.67 (1H, dd), 7.83 (1H,d), 7.36 (1H, d), 7.20 (1H, d), 6.97 (1H, dd), 4.17 (2H, q), 1.35 (3H,t) 1.06 min, 286 [M + H]+.

Intermediate 1422-[(1-methylethyl)oxy]-4-[(5-nitro-2-pyridinyl)oxy]benzonitrile

In a vial 2,4-dihydroxybenzonitrile (300 mg, 2.2 mmol),2-chloro-5-nitropyridine (351.96 mg, 2.22 mmol) and K₂CO₃ (920 mg, 6.62mmol) were dissolved in DMF (5 mL). The reaction was heated for 1 hourunder microwave irradiations (Set Temperature: 110° C.). The reactionmixture was diluted with Et₂O and water, acidified with aqueous 1N HCluntil pH=2, the phases were separated and the organics were dried overNa₂SO₄. The solid was filtered out and the solvent was removed affordingcrude 2-hydroxy-4-[(5-nitro-2-pyridinyl)oxy]benzonitrile (664 mg) as abrown solid. To a solution of this crude in dry DMF (5 mL) potassiumcarbonate (460 mg, 3.33 mmol) and isopropyl bromide (313 μL, 3.33 mmol)were added and the reaction mixture was stirred overnight at 50° C. Thereaction was diluted with brine (10 mL) and extracted with ethyl acetate(2×20 mL). The organic layer was dried (Na₂SO₄), filtered and evaporatedand the residue was purified by flash chromatography on silica gel (SNAP25 g) eluting from 100:0 to 75:25 cyclohexane/ethyl acetate affordingthe title compound (260 mg) as white solid.

¹H-NMR (400 MHz, CDCl₃): δ ppm 9.06 (1H, d), 8.56 (1H, dd), 7.61-7.67(1H, m), 7.15 (1H, d), 6.76-6.84 (2H, m), 4.56-4.68 (1H, m), 1.44 (6H,d).

Intermediate 1434-[(5-amino-2-pyridinyl)oxy]-3-(trifluoromethyl)benzonitrile

To a solution of4-[(5-nitro-2-pyridinyl)oxy]-3-(trifluoromethyl)benzonitrile(Intermediate 120, 83 mg) in THF (3 mL)/water (1.5 mL) was added at roomtemperature, iron (75 mg, 1.34 mmol) and NH₄Cl (72 mg, 1.34 mmol) andthe resulting reaction mixture was stirred overnight. The mixture wasfiltered through a small pad of celite washing with EtOAc and water. Tothe filtered mixture was added an aqueous NaHCO₃ saturated solution andthe two phases were separated. The aqueous phase was extracted withEtOAc and the combined organic phases were dried and evaporated todryness. The crude was purified by flash chromatography (companionsystem, 2×12 g Si cartridge, from 100:0 to 70:30 Cyclohexane/EtOAc) toafford the title compound (72 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 7.97 (1H, s), 7.69-7.79 (2H, m), 7.23(1H, d), 7.16 (1H, dd), 6.93 (1H, d); UPLC_ipqc: 0.91 min, 280 [M+H]+.

The following compounds were prepared using the foregoing methodology,replacing 4-[(5-nitro-2-pyridinyl)oxy]-3-(trifluoromethyl)benzonitrile(Intermediate 120) with the appropriate nitro derivative, as describedin the foregoing Reaction Schemes. Some final products were purified byflash-chromatography (Silica or NH cartridge; Cyclohexane/EtOAc or otherappropriate solvent system). In some cases purification by SCX (MeOH andthen 2M ammonia solution in MeOH) was run before the usualflash-chromatography.

UPLC_ipqc Nitro character- Int. Structure Name derivative NMRcharacterization ization 144

4-[(5-amino-2- pyrimidinyl)oxy]-2- ethylbenzonitrile 2-ethyl-4-[(5-nitro-2- pyrimidinyl)oxy] benzonitrile (Intermediate 133) ¹H-NMR (400MHz, DMSO- d₆): δ ppm 8.03 (2 H, br. s), 7.79 (1H, d), 7.19 (1 H, br.s), 7.07 (1H, d), 5.41 (2H, br. s), 2.80 (2H, q), 1.22 (3H, t) 0.78 min,241 [M + H]⁺ 145

4-[(5-amino-2- pyridinyl)oxy]-3- cyclopropyl- benzonitrile3-cyclopropyl- 4-[(5-nitro-2- pyridinyl)oxy] benzonitrile (Intermediate137) ¹H NMR (400 MHz, DMSO- d₆): δ ppm 7.61-7.52 (2H, m), 7.42 (1H, s),7.16-7.09 (1H, m), 6.89 (2H, t), 5.19 (2H, br. s), 2.14-2.04 (1H, m),0.97-0.89 (2H, m), 0.82- 0.75 (2H, m) 0.86 min, 252 [M + H]⁺ 146

4-[(5-amino-2- pyridinyl)oxy]-3- (1,1- dimethylethyl) benzonitrile3-(1,1- dimethylethyl)- 4-[(5-nitro-2- pyridinyl)oxy] benzonitrile(Intermediate 123) ¹H NMR (400 MHz, DMSO- d₆): δ ppm 7.72 (1H, br. s),7.65-7.57 (2H, m), 7.16- 7.08 (1H, m), 6.89-6.78 (2H, m), 5.28-5.19 (2H,m), 1.39 (9H, s) 1.02 min, 268 [M + H]⁺ 147

6-({4-methyl-3- [(trifluoromethyl) oxy]phenyl}oxy)-3- pyridinamine2-({4-methyl-3- [(trifluoromethyl) oxy]phenyl} oxy)-5- nitropyridine(Intermediate 124) ¹H NMR (400 MHz, CDCl₃): δ ppm 7.70 (1H, d), 7.20(1H, d), 7.09 (1H, dd), 6.95 (1H, br. s.), 6.91 (1H, dd), 6.78 (1H, d),2.27 (3H, s) 1.04 min, 285 [M + H]+ 148

4-[(5-amino-2- pyridinyl)oxy]-2- (methyloxy) benzonitrile 2-(methyloxy)-4-[(5-nitro-2- pyridinyl)oxy] benzonitrile (Intermediate 125) ¹H NMR(400 MHz, DMSO- d₆): δ ppm 7.64 (1H, d) 7.61 (1H, d) 7.11 (1H, dd) 6.87(1H, d) 6.83 (1H, d) 6.52 (1H, dd) 5.27 (2H, br. s.) 3.86 (3H, s) 0.74min, 242 [M + H]+ 149

4-[(4- aminophenyl)oxy]- 2- (methyloxy) benzonitrile 2-(methyloxy)-4-[(4- nitrophenyl)oxy] benzonitrile (Intermediate 126) ¹H NMR (400 MHz,DMSO- d₆): δ ppm 7.63 (1 H, d) 6.81- 6.87 (2 H, m) 6.76 (1 H, d)6.60-6.67 (2 H, m) 6.42 (1 H, dd) 5.12 (2 H, br. s.) 3.86 (3 H, s) 0.70min, 241 [M + H]+ 150

4-[(4- aminophenyl)oxy]- 2- (ethyloxy) benzonitrile 2-(ethyloxy)-4- [(4-nitrophenyl)oxy] benzonitrile (Intermediate 139) ¹H NMR (400 MHz,CDCl₃): δ ppm 7.42 (1H, d) 6.85- 6.90 (2H, m) 6.68-6.75 (2H, m) 6.48(1H, d) 6.44 (1H, dd) 4.04 (2H, q) 3.70 (2H, br. s.) 1.45 (3H, t) 0.80min, 255 [M + H]+ 151

4-[(5-amino-2- pyridinyl)oxy]-2- [(cyclopropylmethyl) oxy]benzonitrile2- [(cyclopropyl- methyl) oxy]-4- [(5-nitro-2- pyridinyl)oxy]benzonitrile (Intermediate 140) ¹H NMR (400 MHz, CDCl₃): δ ppm 7.74 (1H,dd), 7.47 (1H, d), 7.13 (1H, dd), 6.83 (1H, d), 6.61 (1H, d), 6.57 (1H,dd), 3.85 (2H, d), 3.70 (2H, br. s.), 1.21-1.35 (1H, m), 0.60-0.70 (2H,m), 0.32- 0.40 (2H, m) 0.91 min, 282 [M + H]+ 152

4-[(5-amino-2- pyridinyl)oxy]-2- (ethyloxy) benzonitrile 2-(ethyloxy)-4-[(5-nitro-2- pyridinyl)oxy] benzonitrile (Intermediate 141) ¹H NMR (400MHz, CDCl₃): δ ppm 7.75 (1H, dd), 7.48 (1H, d), 7.13 (1H, dd), 6.84 (1H,d), 6.63 (1H, d), 6.58 (1H, dd), 4.08 (2H, q), 3.68 (2H, br. s.), 1.46(3H, t) 0.81 min, 256 [M + H]+ 153

4-[(5-amino-2- pyridinyl)oxy]-2- cyclopropyl- benzonitrile2-cyclopropyl- 4-[(5-nitro-2- pyridinyl)oxy] benzonitrile (Intermediate134) ¹H NMR (400 MHz, DMSO- d₆) δ ppm 7.70 (1 H, d), 7.60 (1 H, d), 7.12(1 H, dd), 6.85 (1 H, d), 6.80 (1 H, dd), 6.65 (1 H, d), 5.26 (2 H, br.s.), 2.11-2.22 (1 H, m), 1.07- 1.15 (2 H, m), 0.75-0.82 (2 H, m) 0.86min, 252 [M + H]+ 154

4-[(5-nitro-2- pyridinyl)oxy]-2-(1- methylethenyl) benzonitrile 2-(1-methylethenyl)- 4-[(5-amino-2- pyridinyl)oxy] benzonitrile (Intermediate138) ¹H-NMR (400 MHz, DMSO- d₆): δ ppm 7.79 (1H, d), 7.62 (1H, d), 7.12(1H, dd), 7.05 (1H, d), 6.98 (1H, dd), 6.89 (1H, d), 5.40 (1H, s), 5.28(2H, br. s.), 5.23 (1H, s), 2.12 (3H, s) 0.90 min, 252 [M + H]+ 155

4-[(5-amino-2- pyridinyl)oxy]-2- ethylbenzonitrile 2-ethyl-4-[(5-nitro-2- pyridinyl)oxy] benzonitrile (Intermediate 135) ¹H-NMR (400 MHz,CDCl₃): δ ppm 7.77 (1H, d) 7.58 (1H, d) 7.15 (1H, dd) 7.00 (1H, d) 6.92(1H, dd) 6.86 (1H, d) 3.62 (2H, br. s.) 2.86 (2H, q) 1.29 (3H, t) 0.86min, 240 [M + H]+ 156

4-[(5-amino-2- pyridinyl)oxy]-2- [(1- methylethyl)oxy] benzonitrile2-[(1- methylethyl)oxy]- 4-[(5-nitro-2- pyridinyl)oxy] benzonitrile(Intermediate 142) ¹H-NMR (400 MHz, DMSO- d₆): δ ppm 7.63 (1H, d), 7.61(1H, d), 7.12 (1H, dd), 6.88 (1H, d), 6.84 (1H, d), 6.51 (1H, dd), 5.29(2H, br. s.), 4.66-4.77 (1H, m), 1.29 (6H, d) 0.89 min, 270 [M + H]+ 157

4-[(5-amino-2- pyridinyl)oxy]-2- ethylbenzonitrile 2-ethyl-4-[(5-nitro-2- pyridinyl)oxy] benzonitrile (Intermediate 135) ¹H NMR (400 MHz,DMSO- d₆): δ ppm 7.73 (1H, d), 7.60 (1H, m), 7.13-7.10 (1H, m), 7.02(1H, d), 6.89-6.86 (2H, m), 5.28 (2H, br. s.), 2.79- 2.73 (2H, q),1.21-1.17 (3H, t) 0.85 min, 240 [M + H]+ 158

4-[(5-amino-2- pyrimidinyl)oxy]-3- ethylbenzonitrile 3-ethyl-4-[(5-nitro-2- pyrimidinyl)oxy] benzonitrile (Intermediate 136) ¹H NMR (400MHz, CDCl₃): δ ppm 8.09 (2H, s), 7.62 (1H, s), 7.55 (1H, dd), 7.15 (1 H,d), 3.62 (2H, br. s.), 2.68 (2 H, q), 1.24 (3 H, t) 0.77 min, 241 [M +H]+ 159

4-[(5-amino-2- pyrimidinyl)oxy]-3- methylbenzonitrile 3-methyl-4-[(5-nitro-2- pyrimidinyl)oxy] benzonitrile (Intermediate 114) ¹H NMR (400MHz, CDCl₃): δ ppm 8.08 (2H, s), 7.60 (1H, s), 7.55 (1H, d), 7.17 (1H,d), 3.59 (2H, br. s.), 2.27 (3H, s) 0.68 min, 227 [M + H]+ 160

4-[(5-amino-2- pyridinyl)oxy]-3- methylbenzonitrile 3-methyl-4-[(5-nitro-2- pyridinyl)oxy] benzonitrile (Intermediate 131) ¹H NMR (400 MHz,CDCl₃): δ ppm 7.71 (1H, d), 7.54 (1H, s), 7.45 (1H, dd), 7.14 (1H, dd),6.95 (1H, d), 6.83 (1 H, d), 3.61 (2H, br. s.), 2.29 (3H, s) 0.75 min,226 [M + H]+ 161

4-[(5-amino-2- pyridinyl)oxy]-2- [(trifluoromethyl) oxy]benzonitrile4-[(5-nitro-2- pyridinyl)oxy]- 2- [(trifluoromethyl) oxy]benzonitrile(Intermediate 132) ¹H NMR (400 MHz, CDCl₃): δ ppm 7.77 (1H, d), 7.66(1H, d), 7.17 (1 H, dd), 7.11 (1 H, s), 7.06 (1 H, dd), 6.89 (1 H, d)0.94 min, 296 [M + H]+

Intermediate 1622-({4-bromo-3-[(trifluoromethyl)oxy]phenyl}oxy)-5-pyrimidinamine

To a solution of 4-bromo-3-[(trifluoromethyl)oxy]phenol (257 mg, 1.0mmol) in dry DMF (4 mL) potassium carbonate (276 mg, 2 mmol) and then2-chloro-5-nitropyrimidine (319 mg, 2.0 mmol) were added and thereaction mixture was stirred for 2 hours at r.t. The reaction wasquenched with water (1 mL), diluted with brine (5 mL) and extracted withethyl acetate (2×15 mL). The organic layer was dried (Na₂SO₄), filteredand evaporated to give crude2-({4-bromo-3-[(trifluoromethyl)oxy]phenyl}oxy)-5-nitropyrimidine. Thiscrude was dissolved in THF/water (2:1) (6 mL) and Iron (279 mg, 5 mmol)and NH₄Cl (267.5 mg, 5 mmol) were added and the reaction mixture wasstirred overnight at r.t. The solid was filtered off and the solutionwas diluted with an aqueous saturated solution of NaHCO₃ (5 mL) andextracted with ethyl acetate (2×20 mL). The organic layer was dried(Na₂SO₄), filtered and evaporated and the residue was purified by flashchromatography on silica gel (SNAP 25 g) eluting from 75:25 to 40:60cyclohexane/ethyl acetate to afford the title compound (280 mg) as lightyellow solid.

¹H-NMR (400 MHz, DMSO-d₆): δ ppm 8.00 (2H, s), 7.81 (1H, d), 7.34-7.40(1H, m), 7.14 (1H, dd), 5.37 (2H, br. s.); UPLC_ipqc: 1.02 min, 350[M]+Br pattern.

The following compounds were prepared using the foregoing methodology,replacing 4-bromo-3-[(trifluoromethyl)oxy]phenol with the appropriatelysubstituted phenol, as described in the foregoing Reaction Schemes.

UPLC_ipqc Int. Structure Name Phenol NMR characterizationcharacterization 163

4-[(5-amino-2- pyrimidinyl)oxy]- 3-(1,1- dimethylethyl) benzonitrile(3-(1,1- dimethylethyl)- 4- hydroxybenzonitrile (Intermediate 109)¹H-NMR (400 MHz, DMSO-d₆): δ ppm 8.01 (2H, s) 7.76 (1H, d) 7.67 (1H, dd)7.05 (1H, d) 5.35- 5.41 (2H, m) 1.35 (9H, s) 0.92 min, 269 [M + H]+

Intermediate 1642-({4-methyl-3-[(trifluoromethyl)oxy]phenyl}oxy)-5-pyrimidinamine

To a solution of2-({4-bromo-3-[(trifluoromethyl)oxy]phenyl}oxy)-5-pyrimidinamine(Intermediate 162, 270 mg) in DMF (4 mL) were added K₃PO₄ (490 mg, 2.31mmol), Pd(tBu₃)₂ (197 mg, 0.385 mmol) and methyl boronic acid (276 mg,4.62 mmol) and the reaction mixture was stirred for 30 minutes at 110°C. under microwave irradiation. After cooling the reaction was dilutedwith water (10 mL) and extracted with ethyl acetate (3×10 mL). Theorganic layer was washed with ice cold brine (2×5 mL), dried (Na₂SO₄),filtered and evaporated and the residue was purified by flashchromatography on silica gel (SNAP 10 g) eluting from 80:20 to 50:50cyclohexane/ethyl acetate to afford the title compound as white solid(135 mg).

UPLC_ipqc: 0.97 min, 286 [M+H]+.

Intermediate 165 methylN-{[(6-{[4-cyano-2-(trifluoromethyl)phenyl]oxy}-3-pyridinyl)amino]carbonyl}-2-methylalaninate

To a solution of triphosgene (32 mg, 0.11 mmol) in EtOAc (1 ml) at 0° C.was added dropwise a solution of4-[(5-amino-2-pyridinyl)oxy]-3-(trifluoromethyl)benzonitrile(Intermediate 143, 69.8 mg) in triethylamine (60 μL)/EtOAc (4 mL) andthen a suspension of methyl 2-methylalaninate hydrochloride(Intermediate 107, 46 mg) in triethylamine (120 μL)/EtOAc (4 mL). Theresulting reaction mixture was stirred for 1 hour. An aqueous pH 3buffer solution was added to the reaction mixture and the two phaseswere separated. The aqueous phase was extracted 3 times with EtOAc andthe combined organic phases were dried over sodium sulphate andevaporated to dryness to give the title compound (49 mg) as crude. Thiscrude was used in the next step without further purification.

UPLC_ipqc: 1.01 min, 423 [M+H]+.

The following compounds were prepared using the foregoing methodology,replacing 4-[(5-amino-2-pyridinyl)oxy]-3-(trifluoromethyl)benzonitrile(Intermediate 143) with the appropriate aniline, as described in theforegoing Reaction Schemes. Final products were isolated as crudes.

UPLC_ipqc Int. Structure Name Aniline characterization 166

methyl N-[({6-[(4- cyano-3- ethylphenyl)oxy]- 3- pyridinyl}amino)carbonyl]-2- methylalaninate 4-[(5-amino-2- pyridinyl)oxy]- 2-ethylbenzonitrile (Intermediate 157) 0.99 min, 383 [M + H]+ 167

methyl N-({[6-({4- cyano-3- [(trifluoromethyl) oxy]phenyl}oxy)-3-pyridinyl]amino} carbonyl)-2- methylalaninate 4-[(5-amino-2-pyridinyl)oxy]- 2- [(trifluoromethyl) oxy]benzonitrile (Intermediate161) 1.05 min, 439 [M + H]+

Intermediate 168 1,1-dimethylethyl{(1R)-1-[({6-[(4-cyano-2-cyclopropylphenyl)oxy]-3-pyridinyl}amino)carbonyl]propyl}carbamate

(2R)-2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)butanoic acid (121.4 mg,0.60 mmol) was dissolved in N,N-Dimethylformamide (1 mL).N,N-Diisopropylethylamine (0.126 mL, 0.72 mmol) andO-(7-Azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (227.2 mg, 0.60 mmol) were added. The reactionmixture was stirred at r.t. for 30 min.4-[(5-amino-2-pyridinyl)oxy]-3-cyclopropylbenzonitrile (Intermediate145, 100 mg) was dissolved in 1.0 mL of DMF and the obtained solutionwas added to the reaction mixture. The reaction mixture was stirred andheated at 60° C. for 2 h. After cooling down to r.t., the reactionmixture was evaporated under vacuum and the crude obtained was chargedon a silica gel column and eluted with Cyclohexane/EtOAc (from 100:0 to50:50 Cyclohexane/EtOAc, then plateau at 50:50) affording 133 mg of thetitle compound.

¹H-NMR (400 MHz, CDCl₃): δ ppm 8.42 (1H, br. s), 8.20-8.10 (2H, m),7.51-7.44 (1H, m), 7.32-7.23 (1H, m), 7.08 (1H, d), 7.03-6.95 (1H, m),4.95 (1H, br. s), 4.16-4.05 (1H, m), 2.07-1.95 (2H, m), 1.77-1.68 (1H,m), 1.47 (9H, s), 1.04 (3H, t), 0.95-0.88 (2H, m), 0.71-0.64 (2H, m);UPLC_ipqc: 1.14 min, 437 [M+H]⁺.

The following compounds were prepared using the foregoing methodology,replacing (2R)-2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)butanoic acidwith the appropriate aminoacid and4-[(5-amino-2-pyridinyl)oxy]-3-cyclopropylbenzonitrile (Intermediate145) with the appropriate aniline, as described in the foregoingReaction Schemes. The reaction was carried out at a suitable temperatureranging from r.t. to high temperature. Final products were purified byflash-chromatography (Silica; Cyclohexane/EtOAc or other appropriatesolvent system).

UPLC_ipqc Int. Structure Name Aminoacid Aniline NMR characterizationcharacterization 169

1,1- dimethylethyl ((1R)-1-{[(6- {[4-cyano-2- (1,1- dimethylethyl)phenyl]oxy}-3- pyridinyl)amino] carbonyl}propyl) carbamate (2R)-2-({[(1,1- dimethylethyl)oxy] carbonyl}amino) butanoic acid 4-[(5-amino-2- pyridinyl) oxy]-3-(1,1- dimethylethyl) benzonitrile(Intermediate 146) ¹H NMR (400 MHz, DMSO-d6): δ ppm 10.20 (1H, br. s),8.38 (1H, br. s), 8.20-8.11 (1H, m), 7.77 (1H, br. s), 7.70- 7.64 (1H,m), 7.15 (1H, d), 7.05 (2H, d), 4.04-3.91 (1H, m), 1.77-1.55 (2H, m),1.41-1.34 (18H, m), 0.93-0.87 (3H, m) 1.25 min, 453 [M + H]⁺. 170

1,1- dimethylethyl ((1R)-1-{[(6- {[4-cyano-3- (methyloxy) phenyl]oxy}-3-pyridinyl)amino] carbonyl}propyl) carbamate (2R)-2- ({[(1,1-dimethylethyl)oxy] carbonyl}amino) butanoic acid 4-[(5- amino-2-pyridinyl) oxy]-2- (methyloxy) benzonitrile (Intermediate 148) ¹H NMR(400 MHz, CDCl₃): δ ppm 8.81 (1H, br. s.) 8.26 (1H, d) 8.09 (1H, br. s.)7.46-7.62 (1H, m) 6.84-7.00 (1H, m) 6.62-6.79 (2H, m) 5.13 (1H, br. s.)4.09- 4.30 (1H, m) 3.88 (3H, s) 1.89-2.04 (1H, m) 1.73- 1.81 (1H, m)1.47 (9H, s) 0.97-1.12 (3H, m) 1.04 min, 427 [M + H]+ 171

1,1- dimethylethyl {2-[(4-{[4- cyano-3- (methyloxy) phenyl]oxy} phenyl)amino]-1,1- dimethyl-2- oxoethyl} carbamate N-{[(1,1- dimethylethyl)oxy]carbonyl}-2- methylalanine 4-[(4- aminophenyl) oxy]-2- (methyloxy)benzonitrile (Intermediate 149) ¹H NMR (400 MHz, CDCl₃): δ ppm 9.02 (1H,br. s.) 7.54-7.61 (2H, m) 7.45 (1H, d) 6.99-7.09 (2H, m) 6.55 (1H, d)6.49 (1H, dd) 4.94 (1H, br. s.) 3.85 (3H, s) 1.59 (6H, s) 1.46 (9H, s)1.11 min, 426 [M + H]+. 172

1,1- dimethylethyl ((1R)-1-{[(4- {[4-cyano-3- (methyloxy) phenyl]oxy}phenyl)amino] carbonyl}propyl) carbamate (2R)-2- ({[(1,1- dimethylethyl)oxy]carbonyl} amino)butanoic acid 4-[(4- aminophenyl) oxy]-2-(methyloxy) benzonitrile (Intermediate 149) ¹H NMR (400 MHz, CDCl₃): δppm 8.66 (1H, br. s.) 7.50-7.59 (2H, m) 7.44 (1H, d) 6.92-7.04 (2H, m)6.54 (1H, d) 6.42 (1H, dd) 5.04-5.25 (1H, m) 4.15-4.27 (1H, m) 3.86 (3H,s) 1.90-2.04 (1H, m) 1.73-1.82 (1H, m) 1.47 (9H, s) 1.04 (3H, t) 1.13min, 426 [M + H]+ 173

1,1- dimethylethyl {2-[(4-{[4- cyano-3- (ethyloxy) phenyl]oxy}phenyl)amino] 1,1- dimethyl-2- oxoethyl} carbamate N-{[(1,1-dimethylethyl) oxy]carbonyl}-2- methylalanine 4-[(4- aminophenyl)oxy]-2- (ethyloxy) benzonitrile (Intermediate 150) ¹H NMR (400 MHz,CDCl₃): δ ppm 9.05 (1H, br. s.) 7.54-7.60 (2H, m) 7.45 (1H, d) 6.97-7.09(2H, m) 6.50-6.54 (1H, m) 6.48 (1H, dd) 4.99 (1H, br. s.) 4.05 (2H, q)1.58 (15H, s) 1.45 (3H, t) 1.17 min, 440 [M + H]+ 174

1,1- dimethylethyl ((1R)-1-{[(4- {[4-cyano-3- (ethyloxy) phenyl]oxy}phenyl)amino] carbonyl}propyl) carbamate (2R)-2- ({[(1,1- dimethylethyl)oxy]carbonyl} amino)butanoic acid 4-[(4- aminophenyl) oxy]-2- (ethyloxy)benzonitrile (Intermediate 150) ¹H NMR (400 MHz, CDCl₃): δ ppm 8.56 (1H,br. s.), 7.51-7.59 (2H, m), 7.44 (1H, d), 6.94- 7.03 (2H, m), 6.52 (1H,d), 6.43 (1H, dd), 5.02-5.18 (1H, m), 4.12-4.23 (1H, m), 4.05 (2H, q),1.90- 2.05 (1H, m), 1.69-1.81 (1H, m), 1.47 (9H, s), 1.46 (3H, t), 1.04(3H, t) 1.19 min, 440 [M + H]+ 175

1,1- dimethylethyl [(1R)-1-({[6- ({4-cyano-3- [(cyclopropylmethyl)oxy]phenyl}oxy)-3- pyridinyl]amino} carbonyl)propyl] carbamate (2R)-2-({[(1,1- dimethylethyl) oxy]carbonyl} amino)butanoic acid 4-[(5-amino-2- pyridinyl) oxy]-2- [(cyclopropylmethyl) oxy]benzonitrile(Intermediate 151) ¹H NMR (400 MHz, CDCl₃): δ ppm 8.86 (1H, br. s.),8.25 (1H, d), 8.00- 8.11 (1H, m), 7.52 (1H, d), 6.84- 6.95 (1H, m), 6.68(1H, d), 6.65 (1H, dd), 5.14 (1H, d), 4.15-4.28 (1H, m), 3.86 (2H, d),1.88- 2.03 (1H, m), 1.73 (1H, s), 1.46 (9H, s), 1.25- 1.35(1H, m), 1.04(3H, t), 0.61-0.70 (2H, m), 0.32- 0.42 (2H, m) 1.17 min, 467 [M + H]+.176

1,1- dimethylethyl [(1R)-1-({[6- ({4-methyl-3- [(trifluoromethyl)oxy]phenyl} oxy)-3- pyridinyl]amino} carbonyl)propyl] carbamate (2R)-2-({[(1,1- dimethylethyl) oxy]carbonyl} amino)butanoic acid 6-({4-methyl-3- [(trifluoromethyl) oxy]phenyl} oxy)-3- pyridinamine(Intermediate 147) ¹H NMR (400 MHz, CDCl₃): δ ppm 8.84 (1H, br. s.),8.19 (1H, d), 7.94- 8.08 (1H, m), 7.22 (1H, d), 6.98-7.02 (1H, m), 6.95(1H, dd), 6.83 (1H, d), 5.22 (1H, d), 4.08-4.33 (1H, m), 2.28 (3H, s),1.87- 2.01 (1H, m), 1.65-1.78 (1H, m), 1.45 (9H, s), 1.03 (3H, t) 1.29min, 470 [M + H]+ 177

1,1- dimethylethyl {(1R)-1-[({6- [(4-cyano-3- cyclopropylphenyl) oxy]-3-pyridinyl}amino) carbonyl]propyl} carbamate (2R)-2- ({[(1,1-dimethylethyl) oxy]carbonyl} amino)butanoic acid 4-[(5- amino-2-pyridinyl) oxy]-2- cyclopropylbenzonitrile (Intermediate 153) ¹H-NMR(400 MHz, CDCl₃): δ ppm 8.54 (1H, br. s.) 8.22 (1H, d) 8.15 (1H, dd)7.60 (1H, d) 6.91- 6.99 (2H, m) 6.68 (1H, d) 4.98 (1H, br. s.) 4.09-4.19 (1H, m) 2.26-2.35 (1H, m) 1.94-2.07 (1H, m) 1.69-1.80 (1H, m) 1.49(9H, s) 1.12-1.19 (2H, m) 1.06 (3H, t) 0.75- 0.82 (2H, m) 1.15 min, 437[M + H]+. 178

1,1- dimethylethyl ((1R)-1-{[(6- {[4-cyano-3-(1- methylethenyl)phenyl]oxy}- 3- pyridinyl)amino] carbonyl}propyl) carbamate (2R)-2-({[(1,1- dimethylethyl) oxy]carbonyl} amino)butanoic acid 4-[(5-amino-2- pyridinyl) oxy]-2-(1- methylethenyl) benzonitrile (Intermediate154) ¹H-NMR (400 MHz, DMSO-d₆): δ ppm 10.21 (1 H, br. s.), 8.39-8.47 (1H, m), 8.17 (1 H, dd), 7.86 (1 H, d), 7.24 (1 H, d), 7.17 (2 H, d),7.01-7.10 (1 H, m), 5.43 (1 H, s), 5.27 (1 H, s), 3.95-4.05 (1 H, m),2.14 (3 H, s), 1.57-1.79 (2 H, m), 1.40 (9 H, s), 0.92 (3 H, t) 1.16min, 437 [M + H]+. 179

1,1- dimethylethyl {(1R)-1-[({6- [(4-cyano-3- ethylphenyl) oxy]-3-pyridinyl}amino) carbonyl]propyl} carbamate (2R)-2- ({[(1,1-dimethylethyl) oxy]carbonyl} amino)butanoic acid 4-[(5- amino-2-pyridinyl) oxy]-2- ethylbenzonitrile (Intermediate 155) ¹H-NMR (400 MHz,DMSO-d₆): δ ppm 10.25 (1H, s) 8.43 (1H, d) 8.16 (1H, dd) 7.80 (1H, d)7.20 (1H, d) 7.15 (1H, d) 7.05- 7.11 (2H, m) 3.94-4.02 (1H, m) 2.79 (2H,q) 1.53- 1.76 (2H, m) 1.39 (9H, s) 1.21 (3H, t) 0.91 (3H, t) 180

1,1- dimethylethyl [(1R)-1-({[2- ({4-methyl-3- [(trifluoromethyl)oxy]phenyl} oxy)-5- pyrimidinyl]amino} carbonyl)propyl] carbamate(2R)-2- ({[(1,1- dimethylethyl) oxy]carbonyl} amino)butanoic acid 2-({4-methyl-3- [(trifluoromethyl) oxy]phenyl} oxy)-5- pyrimidinamine(Intermediate 164) 1.21 min, 471 [M + H]+. 181

1,1- dimethylethyl [(1R)-1-({[6- ({4-cyano-3- [(1- methylethyl)oxy]phenyl}oxy)-3- pyridinyl]amino} carbonyl)propyl] carbamate (2R)-2-({[(1,1- dimethylethyl) oxy]carbonyl} amino)butanoic acid 4-[(5-amino-2- pyridinyl) oxy]-2-[(1- methylethyl) oxy]benzonitrile(Intermediate 156) ¹H-NMR (400 MHz, DMSO-d₆): δ ppm 10.24 (1H, br. s.),8.42 (1H, d), 8.16 (1H, dd), 7.70 (1H, d), 7.15 (1H, d), 7.09(1H, d),7.02 (1H, d), 6.72 (1H, dd), 4.70-4.81 (1H, m), 3.94-4.02 (1H, m), 1.54-1.77 (2H, m), 1.39 (9H, s), 1.30 (6H, d), 0.91 (3H, t) 1.15 min, 455[M + H]+ 182

1,1- dimethylethyl {(1R)-1-[({6- [(4-cyano-2- methylphenyl) oxy]-3-pyridinyl}amino) carbonyl]propyl} carbamate (2R)-2- ({[(1,1-dimethylethyl) oxy]carbonyl} amino)butanoic acid 4-[(5- amino-2-pyridinyl) oxy]-3- methylbenzonitrile (Intermediate 160) ¹H NMR (400MHz, CDCl₃): δ ppm 8.48 (1H, br. s.), 8.12-8.21 (2H, m), 7.59 (1H, s),7.52 (1H, d), 7.09 (1H, d), 6.99 (1H, d), 4.98 (1H, br. s.), 4.07- 4.20(1H, m), 2.26 (3H, s), 1.94-2.09 (1H, m), 1.67- 1.81 (1H, m), 1.42-1.66(9H, m), 1.05 (3H, t) 1.09 min, 411 [M + H]+, 409 [M − H]− 183

1,1- dimethylethyl [(1R)-1-({[6- ({4-cyano-3- [(trifluoromethyl)oxy]phenyl} oxy)-3- pyridinyl]amino} carbonyl)propyl] carbamate (2R)-2-({[(1,1- dimethylethyl) oxy]carbonyl} amino)butanoic acid 4-[(5-amino-2- pyridinyl) oxy]-2- [(trifluoromethyl) oxy]benzonitrile(Intermediate 161) ¹H NMR (400 MHz, CDCl₃): δ ppm 8.64 (1H, br. s.),8.26 (1H, d), 8.15- 8.23 (1H, m), 7.70 (1H, d), 7.20 (1H, s), 7.12-7.18(1H, m), 7.02 (1H, d), 4.93- 5.06 (1H, m), 4.10-4.21 (1H, m), 1.93-2.12(1H, m), 1.67-1.83 (1H, m), 1.49 (9H, s), 1.06 (3H, t) 1.18 min, 481[M + H]+, 479 [M − H]− 184

1,1- dimethylethyl {(1R)-1-[({2- [(4-cyano-2- ethylphenyl) oxy]-5-pyrimidinyl} amino)carbonyl] propyl}carbamate (2R)-2- ({[(1,1-dimethylethyl) oxy]carbonyl} amino)butanoic acid 4-[(5- amino-2-pyrimidinyl) oxy]-3- ethylbenzonitrile (Intermediate 158) ¹H-NMR (400MHz, DMSO-d₆): δ ppm 10.38 (1H, s), 8.84 (2H, s), 7.87 (1H, d), 7.75(1H, dd), 7.34 (1H, d), 7.14 (1H, d), 3.94-4.02 (1H, m), 2.52- 2.58 (2H,m), 1.54-1.76 (2H, m), 1.39 (9H, s), 1.11 (3H, t), 0.91 (3H, t) 185

1,1- dimethylethyl {(1R)-1-[({2- [(4-cyano-2- methylphenyl) oxy]-5-pyrimidinyl} amino)carbonyl] propyl}carbamate (2R)-2- ({[(1,1-dimethylethyl) oxy]carbonyl} amino)butanoic acid 4-[(5- amino-2-pyrimidinyl) oxy]-3- methylbenzonitrile (Intermediate 159) ¹H-NMR (400MHz, DMSO-d₆): δ ppm 10.38 (1H, s), 8.83 (2H, s), 7.87 (1H, d), 7.75(1H, dd), 7.35 (1H, d), 7.14 (1H, d), 3.95-4.01 (1H, m), 2.14 (3H, s),1.54-1.76 (2H, m), 1.39 (9H, s), 0.91 (3H, t) 186

1,1- dimethylethyl ((1R)-1-{[(2- {[4-cyano-2- (1,1- dimethylethyl)phenyl]oxy}-5- pyrimidinyl) amino]carbonyl} propyl)carbamate (2R)-2-({[(1,1- dimethylethyl) oxy]carbonyl} amino)butanoic acid 4-[(5-amino-2- pyrimidinyl) oxy]-3- (1,1- dimethylethyl) benzonitrile(Intermediate 163) 1.17 min, 454 [M + H]+

1,1-dimethylethyl((1R)-1-{[(6-{[4-cyano-3-(1-methylethyl)phenyl]oxy}-3-pyridinyl)amino]carbonyl}propyl)carbamate

To a solution of1,1-dimethylethyl((1R)-1-{[(6-{[4-cyano-3-(1-methylethenyl)phenyl]oxy}-3-pyridinyl)amino]carbonyl}propyl)carbamate(Intermediate 178, 73 mg) in MeOH (10 mL) was added Pd 10% w/w onactivated carbon (14 mg) and the reaction mixture was stirred for 30minutes under H₂ atmosphere (P=1 atm). The catalyst was filtered off andthe solvent removed under reduced pressure. The residue was purified byflash chromatography on silica gel (SNAP 10 g) eluting from 75:25 to40:60 cyclohexane/ethyl acetate affording the title compound (62 mg) aswhite solid.

¹H-NMR (400 MHz, DMSO-d₆): δ ppm 10.24 (1H, br. s.), 8.42 (1H, d), 8.16(1H, dd), 7.78 (1H, d), 7.24 (1H, d), 7.15 (1H, d), 7.07-7.11 (1H, m),7.05 (1H, dd), 3.95-4.02 (1H, m), 3.19-3.27 (1H, m), 1.57-1.76 (2H, m),1.39 (9H, s), 1.26 (6H, d), 0.91 (3H, t); UPLC_ipqc: 1.20 min, 439[M+H]+.

Intermediate 188(2R)-2-amino-N-{6-[(4-cyano-2-cyclopropylphenyl)oxy]-3-pyridinyl}butanamide

1,1-dimethylethyl{(1R)-1-[({6-[(4-cyano-2-cyclopropylphenyl)oxy]-3-pyridinyl}amino)carbonyl]propyl}carbamate(Intermediate 168, 133 mg) was dissolved in DCM (6 mL) and, at 0° C.,TFA (3.0 mL) was slowly added. The reaction mixture was stirred at thattemperature for 2 h. After the removal of the volatiles, the crudeobtained was charged on a SCX cartridge and eluted with MeOH and then 2MNH₃ in MeOH affording 102 mg of the title compound.

¹H-NMR (400 MHz, CDCl₃): δ ppm 9.68 (1H, br. s), 8.32-8.18 (2H, m),7.51-7.43 (1H, m), 7.25-7.31 (1H, m), 7.08 (1H, d), 6.99 (1H, d),3.59-3.51 (1H, m), 2.06-1.95 (2H, m), 1.73-1.63 (1H, m), 1.03 (3H, t),0.95-0.89 (2H, m), 0.74-0.63 (2H, m); UPLC_ipqc: 0.68 min, 337 [M+H]+.

The following compounds were prepared using the foregoing methodology,replacing 1,1-dimethylethyl{(1R)-1-[({6-[(4-cyano-2-cyclopropylphenyl)oxy]-3-pyridinyl}amino)carbonyl]propyl}carbamate(Intermediate 168) with the appropriate N-BOC protected amine, asdescribed in the foregoing Reaction Schemes. Final products werepurified by SCX (MeOH and then 2M ammonia solution in MeOH) andfractions eluted with ammonia, containing the product, were concentratedto provide the free-base. Alternatively, after removing the volatiles,to the crude taken up with an appropriate organic solvent was addedNaHCO₃ saturated aqueous solution, the two phases were separated and theorganic layer was dried, filtered and evaporated affording the finalcompound as the free-base.

UPLC_ipqc N-BOC Protected character- Int. Structure Name amine NMRcharacterization ization 189

(2R)-2-amino-N- (6-{[4-cyano-2- (1,1- dimethylethyl) phenyl]oxy}-3-pyridinyl) butanamide 1,1-dimethylethyl ((1R)-1-{[(6-{[4- cyano-2-(1,1-dimethylethyl) phenyl]oxy}-3- pyridinyl)amino] carbonyl}propyl)carbamate (Intermediate 169) ¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.43 (1H,br. s), 8.25-8.15 (1H, m), 7.78 (1H, br. s), 7.70-7.65 (1H, m), 7.14(1H, d), 7.05 (1H, d), 3.20-3.15 (1H, m), 1.74-1.61 (1H, m), 1.57-1.45(1H, m), 1.36 (9H, s), 0.93 (3H, t) 0.79 min, 353 [M + H]⁺ 190

(2R)-2-amino-N- (6-{[4-cyano-3- (methyloxy) phenyl]oxy}-3- pyridinyl)butanamide 1,1-dimethylethyl ((1R)-1-{[(6-{[4- cyano-3- (methyloxy)phenyl]oxy}-3- pyridinyl)amino] carbonyl}propyl) carbamatae ¹H NMR (400MHz, CDCl₃): δ ppm 9.67 (1H, br. s.) 8.25-8.39 (2H, m) 7.53 (1H, d) 7.00(1H, d) 6.63-6.85 (2H, m) 3.89 (3H, s) 3.40-3.57 (1H, m) 1.94- 2.10 (1H,m) 1.66-1.75 0.61 min, 327 [M + H]+ (Intermediate 170) (1H, m) 1.04 (3H,t) 191

N¹-(4-{[4-cyano- 3-(methyloxy) phenyl]oxy} phenyl)-2- methylalanina-mide 1,1-dimethylethyl {2-[(4-{[4-cyano- 3-(methyloxy) phenyl]oxy}phenyl)amino]- 1,1-dimethyl-2- oxoethyl} ¹H NMR (400 MHz, CDCl₃): δ ppm9.96 (1H, br. s.) 7.62-7.71 (2H, m) 7.45 (1H, d) 6.99-7.09 (2H, m) 6.54(1H, d) 6.49 (1H, dd) 3.85 (3H, s) 1.47 0.65 min, 326 [M + H]+ carbamate(6 H, s) (Intermediate 171) 192

(2R)-2-amino-N- (4-{[4-cyano-3- (methyloxy) phenyl]oxy} phenyl)butanamide 1,1-dimethylethyl ((1R)-1-{[(4-{[4- cyano-3- (methyloxy)phenyl]oxy} phenyl)amino] carbonyl}propyl) carbamate ¹H NMR (400 MHz,CDCl₃): δ ppm 9.59 (1H, br. s.) 7.66 (2H, d) 7.44 (1H, d) 7.04 (2H, d)6.41-6.62 (2H, m) 3.85 (3H, s) 3.41-3.54 (1H, m) 1.92-2.11 (1H, m)1.65-1.79 (1H, m) 0.67 min, 326 [M + H]+ (Intermediate 172) 0.97-1.11(3H, m) 193

N¹(4-{[4-cyano-3- (ethyloxy)phenyl] oxy}phenyl)-2- methylalanina- mide1,1-dimethylethyl {2-[(4-{[4-cyano- 3-(ethyloxy) phenyl]oxy}phenyl)amino]- 1,1-dimethyl-2- ¹H NMR (400 MHz, CDCl₃): δ ppm 9.99 (1H,br. s.) 7.61-7.74 (2H, m) 7.46 (1H, d) 7.01-7.12 (2H, m) 6.53 (1H, d)6.49 (1H, dd) 0.73 min, 340 [M + H]+ oxoethyl} 4.06 (2H, q) 1.49 (6H,carbamate s) 1.47 (3H, t) (Intermediate 173) 194

(2R)-amino-N- (4-{[4-cyano-3- (ethyloxy)phenyl] oxy}phenyl) butanamide1,1-dimethylethyl ((1R)-1-{[(4-{[4- cyano-3- (ethyloxy)phenyl]oxy}phenyl) amino]carbonyl} propyl)carbamate (Intermediate 174) ¹H NMR(400 MHz, CDCl₃): δ ppm 9.61 (1H, br. s.), 7.61-7.69 (2H, m), 7.45 (1H,d), 7.00-7.08 (2H, m), 6.52 (1H, d), 6.47 (1H, dd), 4.05 (2H, q), 3.48(1H, dd), 1.95-2.07 0.74 min, 340 [M + H]+. (1H, m), 1.63-1.69 (1H, m),1.45 (3H, t), 1.04 (3H, t) 195

(2R)-2-amino-N- [6-({4-cyano-3- [(cyclopropyl- methhyl)oxy]phenyl}oxy)-3- pyridinyl] butanamide 1,1-dimethylethyl [(1R)-1-({[6-({4-cyano-3- [(cyclopropyl- methyl)oxy] phenyl}oxy)-3- pyridinyl]amino}carbonyl)propyl] ¹H NMR (400 MHz, CDCl₃): δ ppm 9.70 (1H, br. s.),8.27-8.34 (2H, m), 7.54 (1H, d), 7.00 (1H, d), 6.67-6.72 (2H, m), 3.89(2H, d), 3.50 (1H, dd), 1.96- 2.10 (1H, m), 1.62- 0.73 min, 367 [M + H]+carbamate 1.73 (1H, m), 1.24- (Intermediate 175) 1.38 (1H, m), 1.06 (3H,t), 0.63-0.71 (2H, m), 0.35-0.43 (2H, m) 196

(2R)-2-amino-N- [6-({4-methyl-3- [(trifluoromethyl) oxy]phenyl}oxy)-3-pyridinyl] butanamide 1,1-dimethylethyl [(1R)-1-({[6-({4-methyl-3- [(trifluoromethyl) oxy]phenyl}oxy)- 3-pyridinyl]amino}carbonyl)propyl] carbamate Intermediate 176) ¹H NMR (400 MHz, CDCl₃): δppm 9.61 (1H, br. s.), 8.19-8.26 (2H, m), 7.24 (1H, d), 6.99-7.03 (1H,m), 6.97 (1H, dd), 6.92 (1H, d), 3.43-3.52 (1H, m), 2.29 (3H, s),1.92-2.06 (1H, m), 1.81 (2H, br. 0.83 min, 370 [M + H]+. s.), 1.59-1.73(1H, m), 1.03 (3H, t) 197

(2R)-2-amino-N- {6-[(4-cyano-3- cyclopropyl- phenyl)oxy]-3- pyridinyl}butanamide 1,1-dimethylethyl {(1R)-1-[({6- [(4-cyano-3-cyclopropylphenyl) oxy]-3- pyridinyl}amino) carbonyl]propyl} carbamate(Intermediate 177) ¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.46 (1H, d), 8.21(1H, dd), 7.77 (1H, d), 7.11 (1H, d), 7.00 (1H, dd), 6.81 (1H, d),3.23-3.29 (1H, m), 2.14-2.24 (1H, m), 1.63-1.75 (1H, m), 1.45-1.57 (1H,m), 1.09-1.16 (2H, m), 0.92 (3H, t), 0.79-0.87 0.71 min, 337 [M + H]+(2H, m) 198

(2R)-2-amino-N- (6-{[4-cyano-3- (1-methylethyl) phenyl]oxy}-3-pyridinyl) butanamide 1,1 -dimethylethyl ((1R)-1-{[(6-{[4- cyano-3-(1-methylethyl) phenyl]oxy}-3- pyridinyl)amino] carbonyl}propyl) carbamate(Intermediate 187) ¹H-NMR (400 MHz, DMSO-d₆): δ ppm 8.48 (1H, d) 8.22(1H, dd) 7.79 (1H, d) 7.24 (1H, d) 7.15 (1H, d) 7.05 (1H, dd) 3.19-3.30(2H, m) 1.61-1.74 (1H, m) 1.45-1.56 (1H, m) 1.26 (6H, d) 0.91 (3H, t)0.76 min, 339 [M + H]+ 199

(2R)-2-amino-N- (6-[(4-cyano-3- ethylphenyl)oxy]- 3-pyridinyl}butanamide 1,1-dimethylethyl {(1R)-1-[({6-[(4- cyano-3-ethylphenyl)oxy]- 3- pyridinyl}amino) carbonyl]propyl} carbamate(Intermediate 179) ¹H-NMR (400 MHz, DMSO-d₆): δ ppm 8.48 (1H, d) 8.22(1H, dd) 7.80 (1H, d) 7.19 (1H, d) 7.15 (1H, d) 7.07 (1H, dd) 3.22-3.29(1H, m) 2.79 (2H, q) 1.60-1.74 (1H, m) 1.44- 1.56 (1H, m) 1.21 (3H, t)0.91 (3H, t) 200

(2R)-2-amino-N- [2-({4-methyl-3- [(trifluoromethyl) oxy]phenyl}oxy)-5-pyrimidinyl] butanamide 1,1-dimethylethyl [(1R)-1-({[2-({4- methyl-3-[(trifluoromethyl) oxy]phenyl}oxy)- 5-pyrimidinyl] amino}carbonyl)propyl] carbamate (Intermediate 180) 0.78 min, 371 [M + H]+. 201

(2R)-2-amino-N- [6-({4-cyano-3- [(1- methylethyl)oxy] phenyl}oxy)-3-pyridinyl] butanamide 1,1-dimethylethyl [(1R)-1-({[6-({4- cyano-3-[(1-methylethyl)oxy] phenyl}oxy)-3- pyridinyl]amino} carbonyl)propyl]carbamate (Intermediate 181) ¹H-NMR (400 MHz, CDCl₃): δ ppm 9.70 (1H,br. s.), 8.26-8.38 (2H, m), 7.54 (1H, d), 7.01 (1H, d), 6.74 (1H, d),6.68 (1H, dd), 4.52- 4.66 (1H, m), 3.45- 3.54 (1H, m), 1.97- 2.10 (1H,m), 1.65- 1.76 (1H, m), 1.41 (6H, d), 1.06 (3H, t) 202

(2R)-2-amino-N- {6-[(4-cyano-2- methylphenyl) oxy]-3-pyridinyl}butanamide 1,1-dimethylethyl {(1R)-1-[({6-[(4- cyano-2- methylphenyl)oxy]-3-pyridinyl} amino)carbonyl] propyl} carbamate (Intermediate 182)¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.35- 8.41 (1H, m), 8.15- 8.25 (1H, m),7.84 (1H, s), 7.69 (1H, d), 7.14 (2H, t), 3.23-3.39 (1H, m), 2.16 (3H,s). 1.58- 1.75 (1H, m), 1.43-1.58 (1H, m), 0.90 (3H, t) 0.63 min, 311[M + H]+, 309 [M − H]−. 203

(2R)-2-amino-N- [6-({4-cyano-3- [(trifluoromethyl) oxy]phenyl}oxy)-3-pyridinyl] butanamide 1,1-dimethylethyl [(1R)-1-({[6-({4- cyano-3-[(trifluoromethyl) oxy]phenyl} oxy)-3-pyridinyl] amino} carbonyl)propyl]carbamate (Intermediate 183) ¹H NMR (400 MHz, CDCl₃): δ ppm 9.76 (1H,s), 8.25-8.43 (2H, m), 7.70 (1H, d), 7.20 (1H, s), 7.16 (1H, dd), 7.06(1H, d), 3.44-3.59 (1H, m), 1.53-2.12 (2H, m), 1.07 (3H, t) 0.72 min,381 [M + H]+, 379 [M − H]−. 204

(2R)-2-amino-N- {2-[(4-cyano-2- ethylphenyl) oxy]-5- pyrimidinyl}butanamide 1,1-dimethylethyl {(1R)-1-[({2-[(4- cyano-2-ethylphenyl)oxy]- 5-pyrimidinyl} amino)carbonyl] propyl}carbamate(Intermediate 184) ¹H-NMR (400 MHz, DMSO-d₆): δ ppm 8.90 (2H, s) 7.87(1H, d) 7.75 (1H, dd) 7.34 (1H, d) 4.72 (2H, br. s.) 3.24- 3.30 (1H, m)2.52-2.58 (2H, m) 1.62-1.75 (1H, m) 1.45-1.57 (1H, m) 1.11 (3H, t) 0.91(3H, t) 205

(2R)-2-amino-N- {2-[(4-cyano-2- methylphenyl) oxy]-5- pyrimidinyl}butanamide 1,1-dimethytethyl {(1R)-1-[({2-[(4- cyano-2- methylphenyl)oxy]-5- pyrimidinyl} amino)carbonyl] propyl}carbamate (Intermediate 185)¹H-NMR (400 MHz, DMSO-d₆): δ ppm 8.90 (2H, s) 7.87 (1H, d) 7.75 (1H, dd)7.35 (1H, d) 4.71 (2H, br. s.) 3.23- 3.30 (1H, m) 2.14 (3H, s) 1.62-1.75(1H, m) 1.44-1.57 (1H, m) 0.91 (3H, t) 206

(2R)-2-amino-N- (2-{[4-cyano-2- (1,1- dimethylethyl) phenyl]oxy}-5-pyrimidinyl) butanamide 1,1-dimethylethyl ((1R)-1-{[(2-{[4-cyano-2-(1,1- dimethylethyl) phenyl]oxy)-5- pyrimidinyl) amino]carbonyl}propyl)carbamate (Intermediate 186) 0.73 min, 354 [M + H]+.

Intermediate 2074-({5-[(4R)-4-ethyl-4-methyl-2,5-dioxo-1-imidazolidinyl]-2-pyridinyl}oxy)-2-(1-methylethenyl)benzonitrile

To a solution of triphosgene (118 mg, 0.40 mmol) in dry DCM (3 mL) at 0°C. DIPEA (0.695 ml, 4.0 mmol) was added followed by a solution of4-[(5-amino-2-pyridinyl)oxy]-2-(1-methylethenyl)benzonitrile(Intermediate 154, 100 mg) in dry DCM (6 mL) slowly added (5 minutes).After that (2R)-2-methyl-1-(methyloxy)-1-oxo-2-butanaminium chloride(268 mg, 1.6 mmol) dissolved in dry DCM (3 mL) was added at the sametemperature and the reaction mixture was stirred for 45 minutes at 0° C.The reaction was quenched with water and aqueous buffer (pH 3) was addedwhile the pH was allowed to reach ˜5-6. Ethyl acetate (40 ml) was addedand two phases were separated. The organic layer was washed with brine(2×10 ml), dried (Na2SO4), filtered and evaporated affording the ureaintermediate as yellow foam. This urea was dissolved in MeOH (10 mL),NaOMe (10 mg) was added and the reaction mixture was refluxed for 45minutes under stirring. After cooling the mixture was quenched with anaqueous saturated solution of ammonium chloride (10 mL) and diluted withethyl acetate (20 mL). Two phases were separated and the organic layerwas dried (Na2SO4), filtered and evaporated and the residue was purifiedby flash chromatography on silica gel (SNAP 10 g) eluting from 75:25 to50:50 cyclohexane/ethyl acetate affording the title compound as a whitesolid (100 mg).

¹H NMR (400 MHz, CDCl₃) δ ppm 8.31 (1H, d), 7.87 (1H, dd), 7.71 (1H, d),7.14-7.20 (2H, m), 7.12 (1H, d), 5.94 (1H, br. s.), 5.42 (1H, s), 5.34(1H, s), 2.20 (3H, s), 1.95-2.05 (1H, m), 1.79 (1H, dd), 1.56 (3H, s),1.00 (3H, t); UPLC_ipqc: 1.04 min, 377 [M+H]+.

Example 1(5R)-5-methyl-3-{4-[(3-methylphenyl)oxy]phenyl}-2,4-imidazolidinedione

To Boc-anhydride (0.522 g, 2.394 mmol) in dichloromethane (4 mL) wasadded DMAP (0.248 g, 2.033 mmol) followed by slow addition via syringeof a solution of D-Alanine tertbutyl ester in dichloromethane (4 mL)(prepared from D-Alanine tertbutyl ester HCl salt) (0.410 g) that waspartitioned between DCM and an aqueous solution of Na2CO3. The organiclayer was dried (K2CO3), volatiles were evaporated under vacuum andre-dissolved in dichloromethane. The mixture was stirred for 10 minutesand split into three equal aliquots (solution 1).

To 4-[(3-methylphenyl)oxy]aniline (90 mg, 0.45 mmol) in dichloromethane(1 mL) was added with shaking at 35° C. an aliquot of approximately ⅓ ofsolution 1 slowly via syringe (over ca. 1 min). After 30 min, HCl (ca.0.8 mL) was added and the heterogeneous mixture was heated at 100° C.for 2 hours with shaking, allowing the dichloromethane to distill offthrough a glass capillary. After cooling to room temperature the aqueousHCl was pipetted off and the residue dried under vacuum. The residue waspurified by silica gel chromatography (Biotage instrument, 10 g column)eluting with 0-100% EtOAc/cHex to give after drying the title compoundas a solid (30 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 7.40-7.34 (2H, m), 7.28-7.22 (1H, m),7.12-7.05 (2H, m), 6.98 (1H, d), 6.91-6.84 (2H, m), 5.75 (1H, br. s),4.28 (1H, m), 2.34-2.39 (3H, s), 1.59 (3H, m); LC-MS_A: 2.44 min, 295[M−H]−

Example 2(5R)-5-methyl-3-(4-{[3-(methyloxy)phenyl]oxy}phenyl)-2,4-imidazolidinedione

To a solution of N1-(4-{[3-(methyloxy)phenyl]oxy}phenyl)-D-alaninamide(Intermediate 2, 315 mg) and triethyl amine (0.307 mL, 2.200 mmol) indry ethyl acetate (5 mL), triphosgene (163 mg, 0.550 mmol) was added andthe reaction mixture was stirred for 5 minutes. DMAP (67.2 mg, 0.550mmol) was then added and the reaction mixture was stirred for further 10minutes. The reaction was quenched with a saturated solution of sodiumcarbonate (5 mL), diluted with water (10 mL) and extracted with ethylacetate (3 times 20 mL). The organic layer was dried over sodiumsulphate, filtered and evaporated. The residue was purified by silicagel chromatography (Biotage system, 10 g SNAP column) eluting with agradient cyclohexane/ethyl acetate from 100/0 to 60/40 to afford thetitle compound as a white solid (115 mg).

¹H-NMR (400 MHz, DMSO-d₆) δ ppm: 8.46 (1H, s), 7.27-7.42 (3H, m), 7.09(2H, d), 6.77 (1H, dd), 6.65 (1H, t), 6.60 (1H, dd), 4.26 (1H, m), 3.76(3H, s), 1.36 (3H, d); LC-MS_A: 2.31 min, 311 [M−H]−

Example 3(5R)-3-(4-{[3-(ethyloxy)phenyl]oxy}phenyl)-5-methyl-2,4-imidazolidinedione

The title compound was made in a similar fashion to the preparation ofExample 1 replacing 4-[(3-methylphenyl)oxy]aniline with4-[(3-ethyloxyphenyl)oxy]aniline (Intermediate 4, 0.104 g). After silicagel chromatography a brown gum was obtained and triturated withEt2O:cHex (ca. 1:2, ca. 1.5 mL). This afforded after drying, the titlecompound as a faint brown solid (9 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 7.38 (2H, m), 7.28-7.22 (1H, m),7.14-7.07 (2H, m), 6.73-6.67 (1H, m), 6.66-6.60 (2H, m), 5.56 (1H, br.s), 4.29 (1H, m), 4.02 (2H, q), 1.61-1.57 (3H, m), 1.47-1.39 (3H, m);LC-MS_A: 2.46 min, 325 [M−H]−

Example 4(5R)-3-{4-[(3-chloro-5-fluorophenyl)oxy]phenyl}-5-methyl-2,4-imidazolidinedione

To triphosgene (0.052 g, 0.177 mmol) was added a solution of4-[(3-chloro-5-fluorophenyl)oxy]aniline (Intermediate 6, 0.12 g) intoluene (1 mL) and triethylamine (0.16 g) with shaking. A thick slurryimmediately formed and additional toluene (1 mL) was added. The mixturewas stirred for 3 hours at room temperature and then D-alanine (0.067 g,0.757 mmol) in N,N-dimethylformamide (2 mL) and water (ca. 2 mL) (barelydissolved) was rapidly added via pipette. A two-layer system formed thatwas vigorously stirred for 2 hours and kept overnight at roomtemperature. Volatiles were evaporated, hydrochloric acid (2 mL, 24.35mmol) added and the heterogeneous mixture heated at 100° C. for 2 hours,then allowed to cool to room temperature. After cooling to roomtemperature the aqueous HCl was pipetted off and the residue dried undervacuum. The residue was purified by silica gel chromatography (Biotageinstrument, 10 g column) eluting with 0-100% EtOAc/cHex to give thetitle compound as colourless solid (7 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 7.52-7.41 (2H, m), 7.21-7.11 (2H, m),6.92-6.79 (2H, m), 6.72-6.58 (1H, m), 5.57 (1H, s), 4.31 (1H, dd), 1.61(3H, d); LC-MS_A: 2.58 min, 333 [M−H]−

Example 5(5R)-3-{4-[(3-chloro-4-fluorophenyl)oxy]phenyl}-5-methyl-2,4-imidazolidinedione

A mixture ofN-[({4-[(3-chloro-4-fluorophenyl)oxy]phenyl}amino)carbonyl]-D-alanine(Intermediate 9, 352 mg) in 15 mL of 3N HCl was heated at 100° C. for 16hours. Then sodium carbonate was added and the mixture was adjusted topH=8. The mixture was extracted with ethyl acetate (3 times 50 mL) andthe combined organic phases were washed with brine and dried overmagnesium sulphate. Removal of the solvent afforded a residue which waspurified by column chromatography (MeOH/DCM=1/50) to give 42 mg of thetitle compound as a white solid.

¹HNMR (300 MHz, DMSO): δ ppm 8.47 (1H, s), 7.50-7.45 (1H, t), 7.38-7.36(3H, m), 7.13-7.09 (3H, m), 4.28-4.23 (1H, q), 1.36-1.34 (3H, d);MS_(—)2 (ESI): 335 [M+H]+

Example 6(5S)-3-{4-[(3-chloro-4-fluorophenyl)oxy]phenyl}-5-methyl-2,4-imidazolidinedione

A mixture ofN-[({4-[(3-chloro-4-fluorophenyl)oxy]phenyl}amino)carbonyl]-L-alanine(Intermediate 10, 352 mg) in 15 mL of 3N HCl was heated at 100° C. for16 hours. Then sodium carbonate was added and the mixture was adjustedto pH=8. The mixture was extracted with ethyl acetate (3×50 mL). Thecombined organic phases were washed with brine and dried with magnesiumsulphate. Removal of the solvent afforded crude compound which waspurified by column chromatography (MeOH/DCM=1/50) to give 40 mg of thetitle compound as a white solid.

¹HNMR (300 MHz, CDCl₃): δ ppm 7.41-7.31 (2H, m), 7.15-7.09 (2H, m),7.08-7.03 (2H, m), 6.94-6.90 (1H, m), 5.70 (1H, s), 4.30-4.25 (1H, q),1.59-1.57 (3H, d); MS 1 (ESI): 335 [M+H]+

Example 7(5R)-5-methyl-3-(4-{[2-methyl-5-(methyloxy)phenyl]oxy}phenyl)-2,4-imidazolidinedione

N¹-(4-{[2-methyl-5-(methyloxy)phenyl]oxy}phenyl)-D-alaninamide(Intermediate 16, 118 mg) was dissolved in dry dichloromethane (18 mL).The reaction mixture was cooled down in an ice bath. Triethylamine 0.327mL, 2.350 mmol) was added. Then a solution of triphosgene in drydichloromethane (46.5 mg, 0.157 mmol) dissolved in 7 mL ofdichloromethane was added. The reaction was stirred at 0° C. for 10 minunder argon. A saturated aqueous solution of NaHCO3 was added (18 mL)and the aqueous layer was extracted with dichloromethane 4 times (4×15mL). After drying over sodium sulphate, the solvents were removed undervacuum. The residue was purified by silica gel chromatography (Companionsystem, 12 g cartridge) eluting with a gradient cHex/EOAc from to 100/0to 60/40 during 20 min and 60/40 during 30 min. This afforded the titlecompound (91 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 7.34-7.28 (2H, m), 7.15 (1H, d),7.01-6.92 (2H, m), 6.68 (1H, dd), 6.53 (1H, d), 5.63 (1H, s), 4.25 (1H,dd), 3.74 (3H, s), 2.14 (3H, s), 1.56 (3H, d);

UPLC_B: 0.79 min, 326 [M+1]+

Example 8(5R)-5-methyl-3-(4-{[4-methyl-3-(methyloxy)phenyl]oxy}phenyl)-2,4-imidazolidinedione

To a solution ofN1-(4-{[4-methyl-3-(methyloxy)phenyl]oxy}phenyl)-D-alaninamide(Intermediate 22, 215 mg) in dry dichloromethane (15 mL) triethylamine(0.499 mL, 3.58 mmol) was added and the reaction mixture was cooled to0° C. A solution of triphosgene (96 mg, 0.322 mmol) in drydichloromethane (5 mL) was slowly added and the reaction mixture wasstirred for 30 minutes at the same temperature. The reaction wasquenched with water (10 mL) and extracted with dichloromethane (20 mL).The organic layer was dried over sodium sulphate, filtered andevaporated and the residue was purified by flash chromatography (Biotagesystem, SNAP column) on silica gel using as eluent a gradientcyclohexane/ethyl acetate from 80/20 to 40/60 to afford the titlecompound as a white solid (165 mg).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.45 (1H, s), 7.32 (2H, m), 7.16 (1H,d), 7.05 (2H, m), 6.74 (1H, d), 6.52 (1H, dd), 4.25 (1H, dd), 3.77 (3H,s), 2.14 (3H, s), 1.35 (3H, d); UPLC_B: RT 0.82 min, 327 [M+H]+

Example 9(5R)-5-methyl-3-(6-{[3-(1-methylethyl)phenyl]oxy}-3-pyridinyl)-2,4-imidazolidinedione

N¹-(6-{[3-(1-methylethyl)phenyl]oxy}-3-pyridinyl)-D-alaninamide(Intermediate 26, 35 mg) was dissolved in dry dichloromethane (3 mL).The reaction mixture was cooled down in an ice bath. Triethylamine (98μl, 0.701 mmol) was added. Then a solution of triphosgene in drydichloromethane (13.88 mg, 0.047 mmol dissolved in 1 mL ofdichloromethane) was added dropwise. The reaction mixture was stirred at0° C., under argon, during 10 min. A saturated aqueous solution ofNaHCO3 was added (4 mL) and the aqueous layer was extracted withdichloromethane 4 times (4×5 mL). After drying over sodium sulphate, thesolvents were removed under vacuum. The residue obtained was purified bysilica gel chromatography (Companion system, 2×4 g silica cartridges)with a gradient cyclohexane/ethylacetate from 100/0 to 50/50 during 10min and 50/50 during 20 min. This afforded the title compound as a film(27 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 8.32 (1H, d), 7.78-7.75 (1H, dd), 7.36(1H, t), 7.14 (1H, d), 7.05 (1H, t), 7.01-6.99 (2H, m), 6.55 (1H, s),4.27 (1H, q), 2.96 (1H, m), 1.59 (3H, d), 1.30 (6H, d); UPLC: 0.78 min,326 [M+1]+

Example 10(5R)-5-methyl-3-[6-({3-[(1-methylethyl)oxy]phenyl}oxy)-3-pyridinyl]-2,4-imidazolidinedione

To a solution ofN¹-[6-({3-[(1-methylethyl)oxy]phenyl}oxy)-3-pyridinyl]-D-alaninamide(Intermediate 32, 229 mg) and triethylamine (442 mg, 4.38 mmol) indichloromethane (20 mL) was added a solution of triphosgene (216 mg,0.73 mmol) in dichloromethane (10 mL) dropwise at 0° C. during 5minutes. The resulting mixture was stirred at room temperature for 2hours. The solvent was distilled off and the residue was partitionedbetween dichloromethane (3 times 50 mL) and water (50 mL). The combinedorganic layers were washed with brine (3 times 10 mL), dried over sodiumsulphate, filtered and concentrated to afford a grey solid, which waspurified by silica gel chromatography (PE: EtOAc=2:1) to afford thetitle compound as a white solid (120 mg).

¹H NMR (300 MHz, DMSO-d₆): δ ppm 8.54 (1H, s), 8.15-8.14 (1H, d),7.86-784 (1H, dd), 7.33-7.28 (1H, t), 7.13-7.10 (1H, d), 6.80-6.77 (1H,dd), 6.71-6.67 (2H, m), 4.64-4.58 (1H, m), 4.30-4.25 (1H, q), 1.37-1.35(3H, d), 1.27-1.25 (6H, d).

MS 1 (ESI): 342 [M+H]+

Example 11(5R)-3-{6-[(2,5-dimethylphenyl)oxy]-3-pyridinyl}-5-methyl-2,4-imidazolidinedione

The title compound was made in a similar fashion to the preparation ofExample 9 replacingN1-(6-{[3-(1-methylethyl)phenyl]oxy}-3-pyridinyl)-D-alaninamide withN1-{6-[(2,5-dimethylphenyl)oxy]-3-pyridinyl}-D-alaninamide (Intermediate36, 48 mg) to afford the title compound as a yellow powder (31 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 8.25 (1H, d), 7.74-7.71 (1H, dd), 7.17(1H, d), 6.98-6.88 (3H, m), 6.16 (1H, s), 4.25 (1H, q), 2.32 (3H, s),2.14 (3H, s), 1.56 (3H, d); UPLC: 0.77 min, 312 [M+1]+

Example 12(5R)-3-{6-[(2,3-dimethylphenyl)oxy]-3-pyridinyl}-5-methyl-2,4-imidazolidinedione

N1-{6-[(2,3-dimethylphenyl)oxy]-3-pyridinyl}-D-alaninamide (Intermediate40, 13 mg) was dissolved in dry dichloromethane (2 mL). The reactionmixture (under argon) was cooled down in an ice bath. Triethylamine(38.1 μl, 0.273 mmol) was added. Then a solution of triphosgene in drydichloromethane (5.41 mg, 0.018 mmol, 0.40 equiv dissolved in 1 mL ofDCM) was added dropwise. The reaction mixture was stirred at 0C, underargon, during 10 min. A saturated aqueous solution of NaHCO3 was added(3 mL) and the aqueous layer was extracted with dichloromethane 4 times(4×4 mL). After drying over sodium sulphate, the solvents were removedunder vacuum. The residue was purified by silica gel chromatography(Companion system, 4 g silica cartridge), with a gradientcyclohexane/ethylacetate from 100:0 to 55:45 during 10 min and 55:45during 20 min. This afforded the title compound (9 mg)

¹H NMR (400 MHz, CDCl₃): δ ppm 8.26 (1H, d), 7.75-7.54 (2H, m),7.16-6.92 (3H, m), 5.63 (1H, s), 4.24 (1H, m), 2.34 (3H, s), 2.11 (3H,s), 1.60 (3H, d); UPLC: 0.70 min, 312 [M+H]+

Example 13(5R)-3-{6-[(2,6-dimethylphenyl)oxy]-3-pyridinyl}-5-methyl-2,4-imidazolidinedione

In a 50 mL round-bottomed flaskN¹-{6-[(2,6-dimethylphenyl)oxy]-3-pyridinyl}-D-alaninamide (Intermediate44, 173.8 mg) was dissolved in dichloromethane (5 mL) to give a yellowsolution. The reaction mixture was cooled at 0° C.N,N-dimethyl-4-pyridinamine (36.5 mg, 0.298 mmol), triethylamine (0.208mL, 1.492 mmol) and triphosgene (89 mg, 0.298 mmol) were added. Thereaction mixture was stirred at 0° C. After 20 min the solvent wasevaporated under vacuum to afford a yellow solid. This residue waspurified by silica gel chromatography (Biotage instrument, 10 g SNAPSilica column) eluting with Cyclohexane/EtOAc from 2:1 Cyclohexane/EtOActo 1:3 Cyclohexane/EtOAc in 20 CV; then 1:3 Cyclohexane/EtOAc for 5 CV.The collected fractions afforded the title compound as a colourless oil(162 mg).

¹H NMR (400 MHz, CDCl₃) d ppm 8.26 (1H, d), 7.76 (1H, dd), 7.07-7.19(3H, m), 6.95 (1H, d), 6.67 (1H, br. s), 4.32-4.22 (1H, m), 2.17 (6H,s), 1.57 (3H, d); UPLC_s: 0.80 min, 312 [M+H]+.

Example 14(5R)-3-{6-[(2-ethylphenyl)oxy]-3-pyridinyl}-5-methyl-2,4-imidazolidinedione

In a 50 mL round-bottomed flaskN¹-{6-[(2-ethylphenyl)oxy]-3-pyridinyl}-D-alaninamide (Intermediate 48,170.0 mg) was dissolved in dichloromethane (5 mL) to give a pale yellowsolution. The reaction mixture was cooled at 0° C.N,N-dimethyl-4-pyridinamine (35.7 mg, 0.292 mmol), triethylamine (0.203mL, 1.460 mmol) and triphosgene (87 mg, 0.292 mmol) were added. Thereaction mixture was stirred at 0° C. After 20 min, the reaction mixturewas evaporated in vacuo affording yellow solid that was purified bysilica gel chromatography (Biotage system, 10 g SNAP column) elutingwith a gradient Cyclohexane/EtOAc from 2:1 to 1:3 in 20 CV; then 1:3 for5 CV. The collected fractions afforded the title compound as acolourless oil (153.2 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 8.29 (1H, d), 7.76 (1H, dd), 7.36 (1H,dd), 7.31-7.20 (2H, m), 7.08 (1H, dd), 6.99 (1H, d), 6.68-6.52 (1H, m),4.32-4.23 (1H, m), 2.62 (2H, q), 1.57 (3H, d), 1.26-1.18 (3H, m);UPLC_B: 0.76 mins, 312 [M+H]+.

Example 15(5R)-5-methyl-3-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)-2,4-imidazolidinedione

To a solution ofN1-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)-D-alaninamide(Intermediate 52, 255 mg) in dry dichloromethane (15 mL), TEA (0.590 mL,4.23 mmol) was added and the reaction mixture was cooled to 0° C. Asolution of triphosgene (113 mg, 0.381 mmol) in dry dichloromethane(DCM) (5 mL) was slowly added and the reaction mixture was stirred for30 minutes at the same temperature. The reaction was quenched with water(10 mL) and extracted with dichloromethane (20 mL). The organic layerwas dried over sodium sulphate, filtered and evaporated and the residuewas purified by silica gel chromatography (Biotage system, 10 g SNAPcolumn) using as eluent a gradient cyclohexane/ethyl acetate from 70/30to 30/70. This afforded the title compound as a white solid (172 mg,0.525 mmol).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.54 (1H, br. s), 8.13 (1H, d),7.81-7.86 (1H, m), 7.17 (1H, d), 7.09 (1H, d), 6.79 (1H, d), 6.63 (1H,dd), 4.27 (1H, q), 3.77 (3H, s), 2.15 (3H, s), 1.37 (3H, d); UPLC_B:0.75 min, 328 [M+1]+

Example 16(5R)-5-methyl-3-(6-{[2-methyl-5-(methyloxy)phenyl]oxy}-3-pyridinyl)-2,4-imidazolidinedione

The title compound was made in a similar fashion to the preparation ofExample 7 replacingN¹-(4-{[2-methyl-5-(methyloxy)phenyl]oxy}phenyl)-D-alaninamide withN¹-(6-{[2-methyl-5-(methyloxy)phenyl]oxy}-3-pyridinyl)-D-alaninamide(Intermediate 56, 118 mg) to afford the title compound (78 mg)

¹H NMR (400 MHz, CDCl₃): δ ppm 8.26 (1H, d), 7.73 (1H, dd), 7.17 (1H,d), 6.94 (1H, d), 6.74 (1H, dd), 6.64 (1H, d), 5.88 (1H, s), 4.36-4.14(1H, m), 3.82-3.71 (3H, s), 2.10 (3H, s), 1.57 (3H, d); UPLC_B: 0.71min, 328 [M+1]+

Example 17(5R)-5-methyl-3-(6-{[2-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)-2,4-imidazolidinedione

The title compound was made in a similar fashion to the preparation ofExample 9 replacingN1-(6-{[3-(1-methylethyl)phenyl]oxy}-3-pyridinyl)-D-alaninamide withN¹-(6-{[2-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)-D-alaninamide(Intermediate 63, 200 mg) to afford the title compound (184 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 8.28 (1H, d), 7.77 (1H, m), 7.23 (1H, m),6.98 (1H, d), 6.80-6.73 (2H, m), 6.49 (1H, s), 4.29-4.27 (1H, m), 3.89(3H, s), 2.09 (3H, s), 1.58 (3H, d); UPLC_B: 0.72 min, 328 [M+1]+

Example 18(5R)-5-ethyl-3-(4-{[3-(methyloxy)phenyl]oxy}phenyl)-2,4-imidazolidinedione

To D-2-aminobutyric acid (60.4 mg, 0.586 mmol) in dichloromethane (1 mL)was added DIPEA (0.236 mL, 1.352 mmol) and N-Methyl-N-trifluoroacetamide(269 mg, 1.352 mmol) and the suspension was shaken at 40° C. in a closedvial for 2.5 hours, resulting in a virtually clear solution (Solution1).

To Boc-anhydride (138 mg, 0.631 mmol) in dichloromethane (1 mL) wasadded DMAP (55.1 mg, 0.451 mmol) followed by a solution of4-{[3-(methyloxy)phenyl]oxy}aniline (97 mg, 0.451 mmol) indichloromethane (1 mL). The mixture was stirred for 10 min. The brownsolution thus obtained was added to Solution 1 via syringe with shakingat 35° C. and shaking was continued at this temperature for 2 hours. Thesolution was then kept at room temperature for ca. 64 hours. Conc. aq.HCl (ca. 0.8 mL) was added and the heterogenous mixture was heated at100° C. for 2 hours with shaking, allowing the DCM to distill offthrough a glass capillary. After cooling to room temperature, theresidue was diluted with water (ca. 3 mL) and extracted withdichloromethane (2 times ca. 2 mL). The dichloromethane extracts wereconcentrated under vacuum. The residue was purified by silica gelchromatography (Biotage system, 10 g column) eluting with a gradientcHex/EtOAc from 95/5 to 0/100 to give a brown viscous oil that wasdissolved in Et2O (ca. 0.7 mL) and cHex (ca. 0.1 mL) and kept overnight. The overstanding faint brown solution was decanted from a smallquantity of a brown oil that had separated out. As the solvent wasallowed to evaporate from this solution crystallisation initiated. Thematerial thus obtained was allowed to dry and then triturated with Et2O(2 times ca. 0.5 mL) to afford, after drying, the title compound as asolid (28 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 7.40-7.33 (2H, m), 7.28-7.23 (1H, m),7.15-7.07 (2H, m), 6.75-6.69 (1H, m), 6.67-6.61 (2H, m), 5.64 (1H, br.s), 4.25-4.18 (1H, m), 3.81 (3H, s), 2.10-1.88 (2H, m), 1.09 (3H, t);LC-MS_A: 2.43 min, 325 [M−H]−

Example 19(5R)-5-ethyl-3-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)-2,4-imidazolidinedione

Method A

To a solution of(2R)-2-amino-N-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)butanamide(Intermediate 65, 120 mg) in dry dichloromethane (8 mL) TEA (0.265 mL,1.903 mmol) was added and the reaction mixture was cooled to 0° C. Asolution of triphosgene (50.8 mg, 0.171 mmol) in dry dichloromethane(DCM) (2 mL) was slowly added and the reaction mixture was stirred for30 minutes at the same temperature. The reaction was quenched with water(2 mL) and two phases were separated. The organic layer was dried oversodium sulphate, filtered and evaporated and the residue was purified bysilica gel chromatography (Biotage system, 10 g SNAP column) with aseluent a gradient cyclohexane/ethyl acetate 80/20 to cyclohexane/ethylacetate 50/50 to afford the title compound as a white solid (108 mg).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.61 (1H, s), 8.12 (1H, d), 7.82 (1H,dd), 7.17 (1H, d), 7.08 (1H, d), 6.79 (1H, d), 6.63 (1H, dd), 4.25-4.18(1H, m), 3.77 (3H, s), 2.15 (3H, s), 1.89-1.62 (2H, m), 0.95 (3H, t):UPLC_B: 0.79 min, 342 [M+H]+.

Method B

(2R)-2-amino-N-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)butanamide.2HCl(Intermediate 65b) (750 g) was suspended in dichloromethane (7.5 L) and7.5% sodium carbonate aqueous solution (6 L) and stirred untildissolution. Two phases were separated, the organic one was washed withNaCl 10% aqueous solution (6 L) and concentrated under vacuum at 45° C.to about 3.75 L to remove water via the azeotrope (water ˜0.05%).Dichloromethane was added up to 15 L and then Et₃N (1.35 L) was added.This solution was cooled to 0° C.

Triphosgene (201 g) was dissolved in dichloromethane (4.5 L) and thissolution was added in about 10 minutes to the previous solutionmaintaining the internal temperature at about 10° C. The line was washedwith dichloromethane (375 mL). Work-up: the organic mixture was washedwith a 28% aqueous solution of malic acid (7.5 L) then with a 2% w/waqueous solution of sodium carbonate (7.5 L) and finally with a 20%aqueous solution of NaCl (7.5 L). The organic phase was concentrated tothe lowest volume (about 3.75 L), toluene (2.25 L) was added andconcentrated to low volume (3 L). Toluene (1 L) was further added, andconcentrated to low volume (3 L) to remove all the DCM. A suspension wasobtained and it was stirred for 3 hours then filtered and washed withtoluene (2×1 L). The solid was dried under vacuum at 45° C. untilconstant weight (471 g) of the title compound. The solid wasre-crystallised as follow: 459 g of the title compound was suspended inIsopropanol (1400 mL) and heated until complete dissolution (˜70° C.)then cooled to 20° C., stirred for 3 hours then filtered and washed withIPA (2×700 mL). The solid was dried under vacuum at 45° C. untilconstant weight (412 g) of the title compound. The solid was furthertriturated as follow: 412 g of the title compound was suspended intoluene (1200 mL) at 20° C., stirred for 2 hours then filtered andwashed with toluene (2×420 mL). The solid was dried under vacuum at 45°C. until constant weight (404 g) of the title compound as white solid.

¹³C-NMR (150.81 MHz, DMSO-d₆): δ ppm 173.7, 162.7, 158.6, 155.9, 153.1,145.8, 138.9, 131.1, 124.7, 122.6, 113.0, 111.3, 104.9, 58.0, 55.9,24.9, 16.0, 9.3.

Chiral Chromatography: (Column type: Chiralpack OJ-H 4.6 mm×250 mm, 5μm; Column temperature at 40° C.; Mobile phase: n-Hexane/Ethanol in theratio 55/45% v/v; Flow Rate 0.8 mL/min; detector UV DAD @220 nm) 11.26minutes, enantiomeric excess: 99.58%.

Example 20(5S)-5-ethyl-3-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)-2,4-imidazolidinedione

To a solution of(2S)-2-amino-N-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)butanamide(Intermediate 67, 43 mg) in dry dichloromethane (3 mL), triethylamine(0.095 mL, 0.682 mmol) was added. The reaction mixture was cooled downwith an ice bath. A solution of triphosgene (18.21 mg, 0.061 mmol) indichloromethane (0.750 mL) was then added dropwise and the reactionmixture was stirred during 30 minutes at 0° C. The reaction was quenchedwith water (10 mL). The organic phase was then separated, dried oversodium sulphate, filtered and evaporated. The residue was purified byflash chromatography on silica gel using a column SNAP 10 g andcyclohexane/ethyl acetate from 80/20 40/60 as eluent to afford the titlecompound as a white solid (37.2 mg).

¹H NMR (400 MHz, DMSO): δ ppm 8.60 (1H, s), 8.12 (1H, d), 7.82 (1H, dd),7.16 (1H, d), 7.08 (1H, d), 6.78 (1H, d), 6.63 (1H, dd), 4.21-4.19 (1H,m), 3.76 (3H, s), 2.14 (3H, s), 1.84-1.66 (2H, m), 0.95 (3H, t): UPLC:0.76 min, 342 [M+H]⁺

Example 21(5R)-5-ethyl-3-(6-{[3-(1-methylethyl)phenyl]oxy}-3-pyridinyl)-2,4-imidazolidinedione

(2R)-2-amino-N-(6-{[3-(1-methylethyl)phenyl]oxy}-3-pyridinyl)butanamide(Intermediate 70, 930 mg) was dissolved in dry dichloromethane (100 mL).The reaction mixture, under argon, was cooled down in an ice bath.Triethylamine (2.482 mL, 17.81 mmol) was added. Then a solution oftriphosgene in dry dichloromethane (352 mg, 1.187 mmol dissolved in 40mL of dichloromethane) was added dropwise. The reaction mixture wasstirred at 0° C., under argon, during 10 min. A saturated aqueoussolution of NaHCO3 was added (100 mL) and the aqueous layer wasextracted with dichloromethane 4 times (4×80 mL). After drying oversodium sulphate, the solvent was removed under vacuum. The residueobtained was purified by silica gel chromatography (Companion system,120 g silica cartridge), with a gradient cyclohexane/ethyl acetate 100/0to 55/45. The title compound(5R)-5-ethyl-3-(6-{[3-(1-methylethyl)phenyl]oxy}-3-pyridinyl)-2,4-imidazolidinedionewas obtained as a beige powder (768 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 8.27 (1H, d), 7.74 (1H, dd), 7.32 (1H,t), 7.11 (1H, d), 7.01 (1H, t), 6.98-6.95 (2H, m), 6.40 (1H, s), 4.18(1H, t), 2.96-2.89 (1H, m), 1.99-1.97 (1H, m), 1.93-1.63 (1H, m), 1.27(3H, s), 1.25 (3H, s), 1.05 (3H, t); UPLC: 0.81 min, 340 [M+H]+.

Example 225,5-dimethyl-3-(4-{[3-(methyloxy)phenyl]oxy}phenyl)-2,4-imidazolidinedione

To a solution of2-methyl-N1-(4-{[3-(methyloxy)phenyl]oxy}phenyl)alaninamide(Intermediate 72, 1.33 g) and triethylamine (3.70 mL, 26.6 mmol) in drydichloromethane (80 mL) at 0° C. was added dropwise a solution oftriphosgene (600 mg, 2.022 mmol) in dry dichloromethane (20 mL) and thereaction mixture was stirred for 30 minutes at the same temperature. Thereaction was quenched with a saturated solution of ammonium chloride(100 mL) and two phases were separated. Aqueous layer was extracted withdichloromethane (100 mL) and the two organic phases were collected,dried over sodium sulphate, filtered and evaporated. The residue waspurified by silica gel chromatography (Biotage system, 100 g SNAPcartridge) using as eluent a gradient and cyclohexane/ethyl acetate from80/20 to 40/60 to afford the title compound as a white solid (950 mg).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.54 (1H, br. s), 7.34-7.40 (2H, m),7.31 (1H, t), 7.09 (2H, m), 6.73-6.79 (1H, m), 6.65 (1H, t), 6.56-6.62(1H, m), 3.75 (3H, s), 1.40 (6H, s); UPLC_B-MS: 0.79 min, 327 [M+H]+

The title compound was also obtained with the following alternativeroute. To 2-aminoisobutyric acid (120 mg, 1.164 mmol) in dichloromethane(2 mL) was added DIPEA (0.467 mL, 2.68 mmol) andN-methyl-N-trifluoroacetamide (533 mg, 2.68 mmol) and the suspension wasshaken at 40° C. for 3.5 hours and then at 50° C. in a closed vial for 1hour, resulting in a clear solution (Solution 1).

To Boc2O (406 mg, 1.862 mmol) in dichloromethane (8 mL) was added withcooling in an ice bath DMAP (163 mg, 1.338 mmol) followed by slowaddition (over ca. 5 min) via syringe of a solution of4-{[3-(methyloxy)phenyl]oxy}aniline (288 mg, 1.338 mmol) indichloromethane (2 mL). The mixture was stirred for 15 min at roomtemperature. The brown solution thus obtained was added to Solution 1via syringe with stirring and stirring was continued for 1 hour.Volatiles were evaporated under vacuum.

Concentrated aqueous HCl (ca. 2 mL) was added and the heterogeneousmixture was heated at 100° C. for 2 hours. After cooling to roomtemperature the residue was diluted with water (ca. 10 mL) and extractedwith dichloromethane (3 times ca. 5 mL). The dichloromethane extractswere concentrated under vacuum. The residue was purified by silica gelchromatography (Biotage system, 25 g column) eluting with a gradientcHex/EtOAc from 95/5 to 0/100. Fractions of good purity were collected,triturated with Et2O (2 times ca. 0.5 mL) and dried to give the titlecompound as a solid (40 mg)

Product containing but less pure fractions (by TLC) were combined togive 0.14 g brown material. This was again purified by silica gelchromatography (Biotage system, 95/5 to 40/60) and allowed tocrystallize from Et2O (ca. 2 mL) and washed with some Et2O (2 times ca.0.5 mL). This afforded after drying under vacuum an additional quantityof the title compound (15 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 7.43-7.35 (2H, m), 7.28-7.24 (1H, m),7.11 (2H, d), 6.75-6.69 (1H, m), 6.67-6.61 (2H, m), 5.59-5.66 (1H, br,s), 3.81 (3H, s), 1.61-1.54 (6H, s); UPLC: 0.69 min, 325 [M−H]−

Example 233-{4-[(2,3-dimethylphenyl)oxy]phenyl}-5,5-dimethyl-2,4-imidazolidinedione

N1-{4-[(2,3-dimethylphenyl)oxy]phenyl}-2-methylalaninamide (Intermediate76, 68 mg) was solved in 2 mL of ethyl acetate under nitrogenatmosphere. TEA (0.070 mL, 0.50 mmol) was added followed by a solutionof triphosgene (33.8 mg, 0.11 mmol) in 1.0 mL of ethyl acetate. Afterstirring for 5 minutes, DMAP (13.9 mg, 0.11 mmol) was added and thereaction mixture was stirred at room temperature for 10 minutes. Afterquenching with a saturated solution of NaHCO₃, the mixture was extractedtwo times with ethyl acetate, and the collected organic were dried oversodium sulphate, filtered and evaporated. The residue obtained waspurified by silica gel chromatography eluting with a gradient cHex/EtOAcfrom 100/0 to 0/100. This afforded the title compound (74 mg).

¹H-NMR (400 MHz, DMSO-d₆): δ ppm 8.49 (1H, br. s), 7.38-7.28 (2H, m),7.22-7.05 (2H, m), 6.99-6.87 (2H, m), 6.88-6.81 (1H, m), 2.31 (3H, s),2.10 (3H, s), 1.41 (6H, s); UPLC: 0.74 min, 325 [M+H]+.

Example 243-{6-[(2-ethylphenyl)oxy]-3-pyridinyl}-5,5-dimethyl-2,4-imidazolidinedione

In a 50 mL round-bottomed flaskN¹-{6-[(2-ethylphenyl)oxy]-3-pyridinyl}-2-methylalaninamide(Intermediate 78, 67.1 mg) was dissolved in dichloromethane (5 mL) togive a pale yellow solution. The reaction mixture was cooled at 0° C.N,N-dimethyl-4-pyridinamine (13.28 mg, 0.109 mmol), triethylamine (0.076mL, 0.544 mmol) and triphosgene (32.3 mg, 0.109 mmol) were added. Thereaction mixture was stirred at 0° C. After 20 minutes, the reactionmixture was evaporated under vacuum to afford a yellow solid which waspurified by silica gel chromatography (Biotage system, 10 g SNAP column)eluting with a gradient Cyclohexane/EtOAc from 2:1 to 1:3 in 20 CV; then1:3 for 5 CV. The collected fractions afforded the title compound as awhite solid (60.9 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 8.30 (1H, d), 7.78 (1H, dd), 7.33-7.39(1H, m), 7.20-7.31 (2H, m), 7.06-7.12 (1H, m), 6.98 (1H, d), 6.25 (1H,br. s), 2.62 (2H, q), 1.58 (6H, s), 1.22 (3H, t); UPLC_B: 0.80 min, 326[M+H]+.

Example 253-{6-[(2,6-dimethylphenyl)oxy]-3-pyridinyl}-5,5-dimethyl-2,4-imidazolidinedione

In a 50 mL round-bottomed flaskN¹-{6-[(2,6-dimethylphenyl)oxy]-3-pyridinyl}-2-methylalaninamide(Intermediate 80, 144.4 mg) was dissolved in dichloromethane (5 mL) togive a pale yellow solution. The reaction mixture was cooled at 0° C.N,N-dimethyl-4-pyridinamine (27.7 mg, 0.227 mmol), triethylamine (0.158mL, 1.134 mmol) and triphosgene (67.3 mg, 0.227 mmol) were added. Thereaction mixture was stirred at 0° C. After 20 min, the solvent wasevaporated under vacuum to afford a yellow solid which was purified viasilica gel chromatography (Biotage system, 10 g SNAP column) using aseluent a gradient Cyclohexane/EtOAc from 2:1 to 1:3 in 20 CV; then 1:3for 5 CV. The collected fractions afforded the title compound as a whitesolid (139.7 mg).

¹H NMR (400 MHz, CDCl₃)): δ ppm 8.30-8.26 (1H, m), 7.78 (1H, dd),7.20-7.07 (3H, m), 6.94 (1H, d), 6.33 (1H, br. s), 2.17 (6H, s), 1.58(6H, s): UPLC_s: 0.84 min, 326 [M+H]+.

Example 26(5R)-5-(1-methylethyl)-3-(4-{[4-methyl-3-(methyloxy)phenyl]oxy}phenyl)-2,4-imidazolidinedione

To a solution ofN1-(4-{[4-methyl-3-(methyloxy)phenyl]oxy}phenyl)-D-valinamide(Intermediate 83, 65 mg) in dry dichloromethane (10 mL), TEA (0.138 mL,0.990 mmol) was added and the reaction mixture was cooled to 0° C. Asolution of triphosgene (26.4 mg, 0.089 mmol) in dry dichloromethane (3mL) was slowly added and the reaction mixture was stirred for 1 hour at0° C. The reaction was quenched with water (2 mL) and a saturatedaqueous solution of ammonium chloride (5 mL) and was extracted withdichloromethane (2 times 10 mL). The organic layer was dried over sodiumsulphate, filtered and evaporated and the residue was purified by silicagel flash chromatography (Biotage system, 10 g SNAP column) using aseluent a gradient cyclohexane/ethyl from 100/0 to 60/40 to afford thetitle compound as a white solid (55 mg).

¹H-NMR (400 MHz, DMSO-d₆) δ ppm: 8.52 (1H, s), 7.29 (2H, m), 7.16 (1H,d), 7.05 (2H, m), 6.74 (1H, d), 6.52 (1H, dd), 4.13 (1H, dd), 3.77 (3H,s), 2.14 (4H, s), 1.02 (3H, d), 0.88 (3H, d); UPLC_B: 0.91 min, 355[M+H]+.

Example 27(5R)-5-methyl-3-(2-{[3-(1-methylethyl)phenyl]oxy}-5-pyrimidinyl)-2,4-imidazolidinedione

To a solution ofN¹-(2-{[3-(1-methylethyl)phenyl]oxy}-5-pyrimidinyl)-D-alaninamide(Intermediate 87, 100 mg) in dichloromethane (30 mL) were addedtriphosgene (40 mg, 0.133 mmol) and triethylamine (51 mg, 0.5 mmol). Theresulting mixture was stirred at 0° C. for 30 minutes and water (30 mL)was added. It was extracted with dichloromethane (3 times 30 mL) and thecombined organic layer was dried over sodium sulphate and evaporated toafford the title compound (100 mg).

¹HNMR (400 MHz, CDCl₃): δ ppm 8.62 (2H, s), 7.26-7.30 (1H, t), 7.08-7.05(2H, m), 6.99-6.92 (2H, m), 4.22-4.20 (1H, d), 2.88-2.84 (1H, m),1.47-1.45 (3H, d), 1.19-1.18 (6H, d);

MS_(—)2 (ESI): 326 [M+H]+.

Example 28(5R)-5-ethyl-3-(2-{[3-(ethyloxy)-4-methylphenyl]oxy}-5-pyrimidinyl)-2,4-imidazolidinedione

To a solution of((2R)-2-amino-N-(2-{[3-(ethyloxy)-4-methylphenyl]oxy}-5-pyrimidinyl)butanamide(Intermediate 94, 62 mg) in dry dichloromethane (4 mL), TEA (158 μl,1.13 mmol) was added and the reaction mixture was cooled to 0° C. Asolution of triphosgene (25.2 mg, 0.085 mmol) in dry dichloromethane (2mL) was then added dropwise and the reaction mixture was stirred at 0°C. for 20 minutes. The reaction mixture was evaporated under vacuum. Theresidue obtained was purified by silica gel chromatographychromatography (Companion system, 12 g silica cartridge) withCyclohexane/EtOAc as eluents from 100/0 to 50/50 in 20 minutes and then50/50 during 15 minutes to afford the title compound as a white solid(40 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 8.68 (1H, s), 7.16 (1H, d), 6.82-6.59(2H, m), 6.29 (1H, s), 4.23 (1H, t), 4.12 (1H, q), 2.22 (3H, s), 2.04(2H, s), 2.02-1.95 (1H, m), 1.96-1.80 (1H, m), 1.26 (3H, t), 1.06 (3H,t); UPLC: 0.72 min, 357 [M+H]+

Example 29(5R)-5-(1,1-dimethylethyl)-3-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)-2,4-imidazolidinedione

3-methyl-N¹-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)-D-valinamide(Intermediate 97, 5.2 mg) was dissolved in dry dichloromethane (0.5 mL).The reaction mixture was cooled down in an ice bath. Triethylamine (12.6μl, 0.091 mmol) was added at 0° C. Then 0.5 mL of a solution oftriphosgene in dry dichloromethane (2.47 mg, 0.0083 mmol) was addeddropwise. The reaction mixture was stirred under argon during 20 min at0° C. Some water was added and the aqueous layer was extracted withdichloromethane 4 times. After drying over sodium sulphate, the solventswere removed under vacuum. The residue obtained was purified by flashchromatography on silica gel (Companion system, 4 g silica cartridge)with cyclohexane/ethylacetate as eluents from 100/0 to 60/40 during 15min and 60/40 during 10 min. This afforded the title compound (5.9 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 8.24 (1H, d), 7.69 (1H, dd), 7.17 (1H,d), 7.00 (1H, d), 6.66 (2H, m), 6.46 (1H, br s), 3.89 (1H, s), 3.82 (3H,s), 2.23 (3H, s), 1.13 (9H, s); UPLC_ipqc: 1.11 min, 370 [M+H]+

Example 30(5R)-5-ethyl-5-methyl-3-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)-2,4-imidazolidinedione

To a solution ofN1-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)-D-isovalinamide(Intermediate 99, 42 mg) in dry dichloromethane (6 mL), TEA (0.089 mL,0.638 mmol) was added. The mixture was cooled down to 0° C. and asolution of triphosgene (17.03 mg, 0.057 mmol) in dry dichloromethane(1.500 mL) was added dropwise. The mixture was stirred at thattemperature for 1 hour, then a solution of triphosgene (17.03 mg, 0.057mmol) in dry dichloromethane (DCM) (1.500 mL) was added dropwise again.The reaction was stirred for 30 minutes, it was maintained in theice-bath and quenched with water (10 mL). The mixture was allowed toreach the room temperature then it was extracted with dichloromethane(3×7 mL). The combined organic layers were dried over sodium sulphate,filtered and evaporated. The residue obtained was purified by flashchromatography on silica gel using a 10 g SNAP column andcyclohexane/ethyl acetate as eluents from 80/20 to 50/50 (Biotagesystem). This afforded the title compound as a white solid (24 mg).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.57 (1H, s), 8.13 (1H, d), 7.83 (1H,dd), 7.17 (1H, d), 7.07 (1H, d), 6.79 (1H, d), 6.62 (1H, dd), 3.76 (3H,s), 2.14 (3H, s), 1.57-1.86 (2H, m), 1.39 (3H, s), 0.86 (3H, t); UPLC_B:0.83 min, 354 [M−H]+.

Example 317-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)-5,7-diazaspiro[3.4]octane-6,8-dione

To a solution of1-amino-N-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)cyclobutanecarboxamide(Intermediate 102, 60 mg) in dry dichloromethane (2 mL) TEA (0.128 mL,0.916 mmol) was added. The reaction mixture was cooled in an ice-bath,then a solution of triphosgene (24.47 mg, 0.082 mmol) in drydichloromethane (0.500 mL) was added dropwise. The mixture was stirredat 0° C. for 30 minutes. The reaction was maintained at 0° C. andquenched with water (10 mL). The mixture was extracted withdichloromethane (3×4 mL). The combined organic layers were dried oversodium sulphate, filtered and evaporated. The residue was purified byflash chromatography on silica gel using a 25 g SNAP column and using aseluents cyclohexane/ethylacetate from 100:0 to 50:50. This afforded thetitle compound (52 mg) as a white solid.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.96 (1H, s), 8.14 (1H, d), 7.84 (1H,dd), 7.17 (1H, d), 7.08 (1H, d), 6.79 (1H, d), 6.63 (1H, dd), 3.77 (3H,s), 2.61-2.72 (2H, m), 2.42-2.26 (3H, m), 2.15 (3H, s), 1.82-2.02 (1H,m); UPLC: 0.72 min, 354 [M+H]+.

Example 326-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)-4,6-diazaspiro[2.4]heptane-5,7-dione

To a solution of1-amino-N-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)cyclopropanecarboxamide(Intermediate 105, 58 mg) in dry dichloromethane (2 mL) TEA (0.129 mL,0.925 mmol) was added. The reaction mixture was cooled in an ice-bath,then a solution of triphosgene (24.72 mg, 0.083 mmol) in drydichloromethane (0.500 mL) was added dropwise. The reaction mixture wasstirred at 0° C. for 30 minutes, then the solution of triphosgene (24.72mg, 0.083 mmol) in dry dichloromethane (0.500 mL) was added again. Thereaction mixture was stirred at 0° C. for 30 minutes. The reaction wasmaintained in the ice-bath and quenched with water (10 mL). The organiclayer was separated, dried over sodium sulphate, filtered andevaporated. The residue was purified by flash chromatography on silicagel using a column 25 g SNAP column and cyclohexane/ethylacetate aseluents from 100:0 to 50:50. This afforded the title compound as a whitesolid (20 mg).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.71 (1H, s), 8.18 (1H, d), 7.88 (1H,dd), 7.18 (1H, d), 7.09 (1H, d), 6.79 (1H, d), 6.64 (1H, dd), 3.77 (3H,s), 2.15 (3H, s), 1.20-1.47 (4H, m); UPLC: 0.68 min, 340 [M+H]+.

Example 335,5-dimethyl-3-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)-2,4-imidazolidinedione

Example 34(5R)-5-(1-methylethyl)-3-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)-2,4-imidazolidinedione

Example 353-(6-{[2-(1,1-dimethylethyl)phenyl]oxy}-3-pyridinyl)-5,5-dimethyl-2,4-imidazolidinedione

Example 363-(2-{[2-(1,1-dimethylethyl)phenyl]oxy}-5-pyrimidinyl)-5,5-dimethyl-2,4-imidazolidinedione

Example 37(5R)-5-ethyl-5-methyl-3-(2-{[4-methyl-3-(methyloxy)phenyl]oxy}-5-pyrimidinyl)-2,4-imidazolidinedione

Example 38(5R)-5-ethyl-3-(2-{[3-(ethyloxy)-4-methylphenyl]oxy}-5-pyrimidinyl)-5-methyl-2,4-imidazolidinedione

Example 394-{[5-(4,4-dimethyl-2,5-dioxo-1-imidazolidinyl)-2-pyridinyl]oxy}-2-(1-methylethyl)benzonitrile

4-hydroxy-2-(1-methylethyl)benzonitrile (30 mg) was dissolved in 1 mL ofdimethylformamide.3-(6-fluoro-3-pyridinyl)-5,5-dimethyl-2,4-imidazolidinedione(Intermediate 106, 41.5 mg) and potassium carbonate (51.4 mg, 0.372mmol) were added. The reaction mixture was stirred at 120° C. during 40hours. Some diethylether (4 mL) and water (4 mL) were added. The aqueouslayer was extracted 4 times with diethylether. The gathered organiclayers were dried over sodium sulphate and evaporated giving a residuewhich was purified by mass directed purification (METHOD H). Afterevaporation of the fraction, addition of a solution of saturated NaHCO3(3 mL) and extraction with dichloromethane (four times 4 mL),evaporation afforded the title compound (13 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 8.32 (1H, d), 7.88 (1H, dd), 7.66 (1H,d), 7.17 (1H, dd), 7.08 (2H, m); 5.57 (s, 1H), 3.40 (1H, m), 1.57 (6H,s), 1.33 (6H, d); UPLC_ipqc: 1.01 min, 365 [M+H]+.

Example 405,5-dimethyl-3-[6-({3-[(trifluoromethyl)oxy]phenyl}oxy)-3-pyridinyl]-2,4-imidazolidinedione

3-[(trifluoromethyl)oxy]phenol (19.95 mg, 0.112 mmol) was dissolved in 1mL of dimethylformamide.3-(6-fluoro-3-pyridinyl)-5,5-dimethyl-2,4-imidazolidinedione(Intermediate 106, 25 mg) and potassium carbonate (31.0 mg, 0.224 mmol)were added. The reaction mixture was stirred at 120° C. during 22 hours.Some diethylether (4 mL) and water (4 mL) were added. The aqueous layerwas extracted 4 times with diethylether. The gathered organic layerswere dried over sodium sulphate and evaporated giving a residue, whichwas purified by mass directed purification (METHOD I). After evaporationof the fraction, addition of a solution of saturated NaHCO3 (3 mL) andextraction with dichloromethane (four times 4 mL), evaporation affordedthe title compound as a white solid (23 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 8.30 (1H, d), 7.83 (1H, dd), 7.42 (1H,t), 7.06 (4H, m), 6.36 (s br, 1H), 1.55 (6H, s); UPLC_ipqc: 1.01 min,382 [M+H]+

Example 413-{6-[(4-fluoro-3-methylphenyl)oxy]-3-pyridinyl}-5,5-dimethyl-2,4-imidazolidinedione

The title compound was made in a similar fashion to the preparation ofExample 39 replacing 4-hydroxy-2-(1-methylethyl)benzonitrile with4-fluoro-3-methylphenol (11.30 mg, 0.090 mmol)). For the mass directedpurification METHOD J was used (instead of METHOD H). This afforded thetitle compound as a white solid (17 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 8.26 (1H, d), 7.77 (1H, dd), 7.03-6.92(4H, m), 5.50 (1H, br), 2.29 (3H, s), 1.57 (6H, s); UPLC_ipqc: 0.96 min,330 [M+H]+.

Example 423-{6-[(4-fluoro-2-methylphenyl)oxy]-3-pyridinyl}-5,5-dimethyl-2,4-imidazolidinedione

The title compound was made in a similar fashion to the preparation ofExample 40 replacing 3-[(trifluoromethyl)oxy]phenol with4-fluoro-2-methylphenol (14.13 mg, 0.112 mmol) to afford the titlecompound as a white solid (13 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 8.26 (1H, d), 7.77 (1H, dd), 7.03-6.93(4H, m), 5.50 (1H, br), 2.29 (3H, s), 1.57 (6H, s); UPLC_ipqc: 0.94 min,330 [M+H]+.

Example 434-{[5-(4,4-dimethyl-2,5-dioxo-1-imidazolidinyl)-2-pyridinyl]oxy}-3-ethylbenzonitrile

3-ethyl-4-hydroxybenzonitrile (Intermediate 115, 70 mg) was dissolved in2 mL of dimethylformamide.3-(6-fluoro-3-pyridinyl)-5,5-dimethyl-2,4-imidazolidinedione(Intermediate 106, 102 mg) and potassium carbonate (126 mg, 0.915 mmol)were added. The reaction mixture was stirred at 120° C. during 96 hours.Some diethyl ether (4 mL) and water (4 mL) were added. The aqueous layerwas extracted 4 times with diethyl ether. The gathered organic layerswere dried over sodium sulphate and evaporated giving a residue whichwas purified by flash chromatography on silica gel (companion system, 12g Si cartridge) using as eluent a gradient cyclohexane/ethyl acetate100/0, then from 100/0 to 55/45. Evaporation afforded the title compound(54 mg) as a white solid.

¹H NMR (400 MHz, CDCl₃): δ ppm 8.27 (1H, d), 7.87 (1H, dd), 7.62 (1H,d), 7.55 (1H, dd), 7.16 (1H, dd); 6.40 (1H, s), 5.58 (1H, s), 2.65 (2H,q), 1.56 (6H, s), 1.21 (3H, t); UPLC_ipqc: 0.97 min, 349 [M−H]−.

The following compounds were prepared using the foregoing methodology,reacting 3-(6-fluoro-3-pyridinyl)-5,5-dimethyl-2,4-imidazolidinedione(Intermediate 106) with the appropriately substituted phenol for a timeranging from 12 to 96 hrs, as described in the foregoing ReactionSchemes. Some final products were purified by flash-chromatography(Silica or NH cartridge; Cyclohexane/EtOAc or other appropriate solventsystem) and were isolated as the free-base; alternatively, some productswere purified by mass directed purification (METHOD K ChromatographicAcidic conditions: Column=Waters Sunfire OBD (150 mm×30 mm, 5 μmparticle size) at room temperature; Mobile phase=A (water+0.1% formicacid in water), B (acetonitrile+0.1% formic acid in acetonitrile); Flowrate=40 mL/min; Gradient=from 1% (B) to 100% (B) in 8.5 min, 100% (B)during 6.5 min, return to 1% (B) in 0.5 min) and fractions containingthe product were basified with NaHCO₃ and extracted with an appropriateorganic solvent, dried and concentrated to provide the free-base.Finally in one case (Example 77) an additional purification by SCX (DCMand MeOH as solvents) was run.

UPLC_ipqc characteri- Ex. Structure Name Phenol NMR characterizationzation 44

2-chloro-4-{[5- (4,4-dimethyl-2,5- dioxo-1- imidazolidinyl)-2-pyridinyl]oxy} benzonitrile 2-chloro-4 hydroxybenzo- nitrile ¹H NMR (400MHz, CDCl₃): δ ppm 8.35 (1H, d), 7.92 (1H, dd), 7.71 (1H, d), 7.38 (1H,d), 7.21 (1H, dd); 7.14 (1H, d), 5.58 (s, 1H), 1.59 (6H, s) 1.94 min,357 [M + H]+ 45

5,5-dimethyl-3-[6- ({4-methyl-3- [(trifluoromethyl) oxy]phenyl}oxy)- 3-pyridinyl]-2,4- imidazolidinedione 4-methyl-3- [(trifluoro- methyl)oxy]phenol (Intermediate 116) ¹H NMR(400 MHz, CDCl₃): δ ppm 8.28 (1H,d), 7.80 (1H, dd), 7.23-7.34 (1H, m), 7.00-7.09 (2H, m), 6.19 (1H, br.s.), 2.32 (3H, s), 1.56 (6H, s) 1.13 min, 396 [M + H]+. 46

4-{[5-(4,4- dimethyl-2,5- dioxo-1- imidazolidinyl)-2- pyridinyl]oxy}-2-(methyloxy) benzonitrile 4-hydroxy-2- (methyloxy) benzonitrile ¹H NMR(400 MHz, CDCl₃): δ ppm 8.33 (1H, d) 7.88 (1H, dd) 7.58 (1H, d) 7.10(1H, d) 6.76- 6.83 (2H, m) 5.92 (1H, br. s.) 3.92 (3H, s) 1.58 (6H, s)0.86 min, 353 [M + H]+ 47

4-{[5-(4,4- dimethyl-2,5- dioxo-1- imidazolidinyl)-2- pyridinyl]oxy}-3-methylbenzonitrile 4-hydroxy-3- methylbenzo- nitrile (Intermediate 118)¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.66 (1H, s), 8.11-8.19 (1H, m),7.91-8.00 (1H, m), 7.87 (1H, s), 7.76 (1H, d), 7.29 (2H, t), 2.17 (3H,s), 1.41 (6H, s) 0.89 min, 337 [M + H]+, 335 [M − H]−

Example 484-{[5-(4,4-dimethyl-2,5-dioxo-1-imidazolidinyl)-2-pyridinyl]oxy}-3-(trifluoromethyl)benzonitrile

A solution of methylN-{[(6-{[4-cyano-2-(trifluoromethyl)phenyl]oxy}-3-pyridinyl)amino]carbonyl}-2-methylalaninate(Intermediate 165, 49 mg) in MeOH (10 mL) was heated at reflux for 10min. Sodium methoxide (4 mg) was added to the hot reaction mixture andit was left refluxing for 2.5 hours. The solvent was evaporated todryness and the residue obtained was purified by flash-chromatography(companion system, 12 g Si cartridge), with Cyclohexane/EtOAc as eluentsfrom 70/30 to 50/50, to give two batches. The second batch was purifiedagain by mass directed purification (METHOD K), leading to the desiredcompound which was gathered to the first batch, to afford the titlecompound as a white solid (8.2 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 8.30 (1H, d), 8.02 (1H, s), 7.90-7.99(1H, m), 7.87 (1H, d), 7.46 (1H, d), 7.21 (1H, d), 5.68 (1H, br. s.),1.59 (6H, s); UPLC_ipqc: 0.98 min, 391 [M+H]+.

The following compounds were prepared using the foregoing methodology,replacing methylN-{[(6-{[4-cyano-2-(trifluoromethyl)phenyl]oxy}-3-pyridinyl)amino]carbonyl}-2-methylalaninate(Intermediate 165) with the appropriate urea, as described in theforegoing Reaction Schemes. Final products were purified byflash-chromatography (Silica cartridge; Cyclohexane/EtOAc or otherappropriate solvent system) and were isolated as the free-base.

UPLC_ipqc Ex. Structure Name Urea NMR characterization characterization49

4-{(5-(4,4-dimethyl- 2,5-dioxo-1- imidazolidinyl)-2- pyridinyl]oxy}-2-ethylbenzonitrile methyl N-[({6-[(4- cyano-3- ethylphenyl)oxy]-3-pyridinyl}amino) carbonyl]-2- methylalaninate (Intermediate 166) ¹ HNMR(400 MHz, CDCl₃): δ ppm 8.32 (1H, d), 7.84-7.90 (1H, m), 7.65 (1H,d), 7.04-7.16 (3H, m), 5.49 (1H, s), 2.90 (2H, q), 1.57-1.59 (6H, s),1.32 (3H, t) 0.97 min, 351 [M + H]+. 50

4-{[5-(4,4-dimethyl- 2,5-dioxo-1- imidazolidinyl)-2- pyridinyl]oxy}-2-[(trifluoromethyl) oxy]benzonitrile methyl N-({[6-({4- cyano-3-[(trifluoromethyl) oxy]phenyl}oxy)-3- pyridinyl]amino} carbonyl)-2-methylalaninate (Intermediate 167) ¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.66(1H, s), 8.21-8.28 (1H, m), 8.13 (1H, d), 7.97- 8.05 (1H, m), 7.64 (1H,s), 7.41-7.49 (1H, m), 7.35 (1H, d), 1.41 (6H, s) 1.01 min, 407 [M +H]+, 405 [M − H]−.

Example 514-{[5-(4,4-dimethyl-2,5-dioxo-1-imidazolidinyl)-2-pyrimidinyl]oxy}-2-ethylbenzonitrile

To a solution of triphosgene (20.8 mg, 0.07 mmol) in EtOAc (0.5 mL) at0° C., a solution of 4-[(5-amino-2-pyrimidinyl)oxy]-2-ethylbenzonitrile(Intermediate 144, 35 mg)/TEA (0.037 mL, 0.27 mmol) in EtOAc (1.0 mL)was added dropwise under stirring. Then a solution of methyl2-methylalaninate hydrochloride (Intermediate 107, 33.6 mg)/TEA (0.073mL, 0.52 mmol) in 1.0 mL of EtOAc at 0° C. was slowly added and thereaction mixture was stirred at that temperature for 10 minutes. Themixture was quenched with an aqueous pH 3 buffer solution to pH 5-6 andextracted with EtOAc (three times). The collected organic were driedover Na₂SO₄, filtered and evaporated. The crude obtained was dissolvedin MeOH (1.0 mL) and 3.0 mg (0.056 mmol) of sodium methoxide were addedand the mixture was heated at 60° C. for 5 minutes. After cooling downto r.t., the mixture was evaporated, diluted with EtOAc and washed withNH₄Cl (aqueous saturated solution) and the aqueous phase was extractedwith EtOAc (three times). The collected organic were dried over Na₂SO₄,filtered and evaporated. The crude obtained was charged on a silica gelcolumn and eluted with Cyclohexane/EtOAc (from 70:30 Cyclohexane/EtOActo 60:40 in 20 CV, plateau at 60:40 in 20 CV) affording 11.5 mg of thetitle compound.

¹H-NMR (400 MHz, CDCl₃): δ ppm 8.77 (2H, br. s), 7.69 (1H, d), 7.21 (1H,br. s), 7.16 (1H, d), 5.89 (1H, br. s), 2.91 (2H, q), 1.60 (6H, s), 1.33(3H, t); UPLC_ipqc: 0.92 min, 352 [M+H]⁺.

The following compounds were prepared using the foregoing methodology,replacing 4-[(5-amino-2-pyrimidinyl)oxy]-2-ethylbenzonitrile(Intermediate 144) with the appropriate aniline, as described in theforegoing Reaction Schemes. Final products were purified byflash-chromatography (Silica or NH cartridge; Cyclohexane/EtOAc,dichlorometane/methanol or other appropriate solvent system);alternatively, some products were purified by mass directed purificationand fractions containing the product were basified with NaHCO₃ andextracted with an appropriate organic solvent, dried and concentrated toprovide the desired product.

UPLC_ipqc characteri- Ex. Structure Name Aniline NMR characterizationzation 52

3-cyclopropyl-4- {[5-(4,4-dimethyl- 2,5-dioxo-1- imidazolidinyl)-2-pyridinyl]oxy} benzonitrile 4-[(5-amino-2- pyridinyl)oxy]-3-cyclopropylbenzo- nitrile (Intermediate 145) ¹H NMR (400 MHz, DMSO-d₆):δ ppm 8.65 (1H, br. s), 8.16 (1H, br. s), 7.99-7.91 (1H, m), 7.73-7.66(1H, m), 7.52 (1H, br. s), 7.29 (2H, d), 1.97-1.86 (1H, m), 1.41 (6H,s). 0.91- 0.83 (2H, m), 0.80-0.74 (2H, m) 0.97 min, 363 [M + H]⁺ 53

4-{[5-(4,4-dimethyl- 2,5-dioxo-1- imidazolidinyl)-2- pyridinyl]oxy}-3-(1,1- dimethylethyl) benzonitrile 4-[(5-amino- 2-pyridinyl)oxy]-3-(1,1-dimethylethyl) benzonitrile (Intermediate 146) ¹H NMR (400 MHz,DMSO-d₆): δ ppm 8.66 (1H, br. s), 8.25- 8.19 (1H, m). 8.02-7.95 (1H, m),7.83 (1H, br. s), 7.76- 7.70 (1H, m), 7.31 (1H, d), 7.19 (1H, d), 1.42(6H, s), 1.36 (9H, m) 1.08 min, 379 [M + H]⁺ 54

2-[{cyclopropyl- methyl)oxy]-4- {[5-(4,4- dimethyl-2,5- dioxo-1-imidazolidinyl)-2- pyridinyl]oxy} benzonitrile 4-[(5-amino-2-pyridinyl)oxy]-2- [(cyclopropyl methyl)oxy] benzonitrile (Intermediate151) ¹H NMR (400 MHz, CDCl₃): δ ppm 8.32 (1H, d). 7.86 (1H, dd), 7.58(1H, d), 7.09 (1H, d), 6.74-6.79 (2H, m), 6.00 (1H, br. s.), 3.91 (2H,d), 1.58 (6H, s), 1.29-1.39 (1H, m), 0.63- 0.72 (2H, m), 0.36-0.44 (2H,m) 1.01 min, 393 [M + H]+ 55

4-{[5-(4,4-dimethyl- 2,5-dioxo-1- imidazolidinyl)-2- pyridinyl]oxy}-2-(ethyloxy) benzonitrile 4-[(5-amino-2- pyridinyl)oxy]-2- (ethyloxy)benzonitrile (Intermediate 152) ¹H NMR (400 MHz, CDCl₃): δ ppm 8.32 (1H,dd), 7.87 (1H, dd), 7.58 (1H, d), 7.09 (1H, d), 6.74-6.80 (2H, m), 5.88(1H, br. s.), 4.12 (2H, q), 1.58 (6H, s), 1.49 (3H, t) 0.93 min, 367[M + H]+ 56

2-cyclopropyl-4- {[5-(4,4-dimethyl- 2,5-dioxo-1- imidazolidinyl)-2-pyridinyl]oxy} benzonitrile 4-[(5-amino-2- pyridinyl)oxy]-2-cyclopropylbenzo- nitrile (Intermediate 153) ¹H NMR (400 MHz, DMSO-d₆):δ ppm 8.64 (1H, s). 8.22 (1H, d), 7.96 (1H, dd), 7.82 (1H, d), 7.25 (1H,d), 7.13 (1H, dd), 6.93 (1H, d), 2.16-2.26 (1H, m), 1.43 (6H, s),1.09-1.18 (2H, m), 0.86 (2H, dd) 0.98 min, 363 [M + H]+ 57

5,5-dimethyl-3-[2- ({4-methyl-3- [(trifluoromethyl) oxy]phenyl}oxy)-5-pyrimidinyl]-2,4- imidazolidinedione 2-({4-methyl-3- [(trifluoromethyl)oxy]phenyl}oxy)-5- pyrimidinamine (Intermediate 164) ¹H-NMR (400 MHz,CDCl₃): δ ppm 8.76 (2H, s), 7.33 (1H, d), 7.12-7.16 (1H, m), 7.12 (1H,dd), 4.68 (1H, br. s.), 2.36 (3H, s), 1.61 (6H, s). 1.08 min, 397 [M +H]+. 58

4-{[5-(4,4-dimethyl- 2,5-dioxo-1- imidazolidinyl)-2- pyrimidinyl]oxy}-3-(1,1- dimethylethyl) benzonitrile 4-[(5-amino- 2- pyrimidinyl)oxy]-3-(1,1- dimethylethyl) benzonitrile (Intermediate 163) ¹H-NMR (400MHz, DMSO- d₆): δ ppm 8.80 (2H, s) 7.79 (1H, d) 7.58 (1H, dd) 7.16 (1H,d) 5.59 (1H, br. s.) 1.62 (6H, s) 1.41 (9H, s) 1.03 min, 380 [M + H]+.59

4-{[5-(4,4-dimethyl- 2,5-dioxo-1- imidazolidinyl)-2- pyridinyl]oxy}-2-[(1- methylethyl)oxy] benzonitrile 4-[(5-amino-2- pyridinyl)oxy]-2-[(1-methylethyl) oxy]benzonitrile (Intermediate 156) ¹H-NMR (400 MHz,DMSO- d₆): δ ppm 8.66 (1H, s), 8.23 (1H, d), 7.96 (1H, dd), 7.76 (1H,d), 7.27 (1H, d), 7.16 (1H, d), 6.83 (1H, dd), 4.74-4.85 (1H, m), 1.42(6H, s), 1.31 (6H, d) 0.98 min, 381 [M + H]+

Example 604-({5-[(4R)-4-ethyl-4-methyl-2,5-dioxo-1-imidazolidinyl]-2-pyridinyl}oxy)-2-[(1-methylethyl)oxy]benzonitrile

In a 2-necked round bottomed flask, bis(trichloromethyl)carbonate (174.5mg, 0.59 mmol) was dissolved in DCM (10 mL). N,N-diethylethanamine(0.651 mL, 3.74 mmol) was added therein. The obtained solution wascooled to 0° C. In a separate vial,4-[(5-amino-2-pyridinyl)oxy]-2-[(1-methylethyl)oxy]benzonitrile(Intermediate 156, 145.4 mg) was dissolved in DCM (10 mL). The obtainedsolution was added dropwise in 5 min to the chilledbis(trichloromethyl)carbonate solution. At the end of addition, at 0°C., having checked that4-[(5-amino-2-pyridinyl)oxy]-2-[(1-methylethyl)oxy]benzonitrilecompletely disappeared, a solution in DCM (5 mL) ofN,N-diethylethanamine (0.280 mL, 1.60 mmol) and(2R)-2-methyl-1-(methyloxy)-1-oxo-2-butanaminium chloride (Tetrahedron1988, 44(15), 4793-6, 179.2 mg, 1.07 mmol) was added to the reactionmixture and it was stirred for 25 min. The reaction mixture was quenchedwith 10 mL of water, diluted with 50 mL of DCM, and acidified to pH ˜5-6using an aqueous pH 3 buffer solution. Phases were separated. Theorganic layer was washed with 15 mL of brine, dried over dry sodiumsulphate, filtered and evaporated in vacuo to obtain a white foam. Thisfoam was dissolved in 8 mL of methanol and Sodium methoxide (17.3 mg,0.32 mmol) was added to the reaction mixture. The vial was sealed andshaken on PLS at 60° C. After 15 min, further sodium methoxide (17.3 mg,0.32 mmol) was added portionwise to the reaction mixture and it wasshaken up to 1.5 hr. The reaction mixture was then evaporated in vacuousing a biotage V10 apparatus to obtain a pale yellow solid. This crudewas purified by Biotage SP1 (Silica; from 100:0 to 50:50Cyclohexane/EtOAc in 10 CV) to give the title compound (116.5 mg).

¹H NMR (400 MHz, CDCl₃) δ ppm 8.30 (1H, d), 7.84 (1H, dd), 7.56 (1H, d),7.08 (1H, d), 6.78 (1H, d), 6.75 (1H, dd), 6.10 (1H, br. s.), 4.49-4.69(1H, m), 1.91-2.04 (1H, m), 1.71-1.83 (1H, m), 1.55 (3H, s), 1.41 (3H,s), 1.40 (3H, s), 0.98 (3H, t); UPLC_ipqc: 1.03 min, 395 [M+H]+.

The following compounds were prepared using the foregoing methodology,replacing4-[(5-amino-2-pyridinyl)oxy]-2-[(1-methylethyl)oxy]benzonitrile(Intermediate 156) with the appropriate aniline, as described in theforegoing Reaction Schemes.

UPLC_ipqc characteri- Ex. Structure Name Aniline NMR characterizationzation 61

3-cyclopropyl-4- ({5-[(4R)-4-ethyl-4- methyl-2,5-dioxo-1-imidazolidinyl]-2- pyridinyl}oxy) benzonitrile 4-[(5-amino-2-pyridinyl)oxy]- 3-cyclopropyl- benzonitrile (Intermediate 145) ¹H NMR(400 MHz, CDCl₃) δ ppm 8.27 (1H, d), 7.85 (1H, dd), 7.52 (1H, dd), 7.30(1H, d), 7.17 (1H, d), 7.11 (1H, d), 5.41 (1H, br. s.), 1.95-2.07 (2H,m), 1.73-1.84 (1H, m), 1.57 (3H, s), 1.00 (3H, t), 0.92- 0.98 (2H, m),0.68-0.75 (2H, m) 1.02 min, 377 [M + H]+. 62

4-({5-[(4R)-4-ethyl- 4-methyl-2,5- dioxo-1- imidazolidinyl]-2-pyridinyl}oxy)-2- [(trifluoromethyl) oxy]benzonitrile 4-[(5-amino-2-pyridinyl)oxy]- 2-[(trifluoro- methyl) oxy]benzonitrile (Intermediate161) ¹H NMR (400 MHz, CDCl₃) δ ppm 8.34 (1H, d), 7.92 (1H, dd), 7.75(1H, d), 7.29-7.32 (1H, m), 7.25 (1H, dd), 7.16 (1H, d), 5.49 (1H, br.s.), 1.96-2.08 (1H, m), 1.74-1.85 (1H, m), 1.58 (3H, s), 1.01 (3H, t)1.06 min, 421 [M + H]+. 63

2-cyclopropyl-4- ({5-[(4R)-4-ethyl-4- methyl-2,5-dioxo-1-imidazolidinyl]-2- pyridinyl}oxy) benzonitrile 4-[(5-amino-2-pyridinyl)oxy]- 2-cyclopropyl- benzonitrile (Intermediate 153) ¹H NMR(400 MHz, CDCl₃) δ ppm 8.29 (1H, dd), 7.84 (1H, dd), 7.64 (1H, d), 7.08(1H, d), 7.02 (1H, dd), 6.75 (1H, d), 5.47 (1H, br. s.), 2.28-2.37 (1H,m), 1.96-2.07 (1H, m), 1.73-1.84 (1H, m), 1.57 (3H, s), 1.14-1.21 (2H,m), 1.00 (3H, t), 0.79-0.85 (2H, m) 1.02 min, 377 [M + H]+. 64

(5R)-5-ethyl-5- methyl-3-[2-({4- methyl-3- [(trifluoromethyl)oxy]phenyl}oxy)-5- pyrimidinyl]-2,4- imidazolidinedione 2-({4-methyl-3-[(trifluoro- methyl)oxy] phenyl}oxy)-5- pyrimidinamine (Intermediate164) ¹H NMR (400 MHz, CDCl₃) δ ppm 8.73 (2H, s), 7.32 (1H, d) 7.12-7.16(1H, m), 7.11 (1H, dd), 5.51 (1H, br. s.), 2.35 (3H, s), 1.95-2.07 (1H,m), 1.74-1.85 (1H, m), 1.00 (3H, t) 1.12 min, 411 [M + H]+. 65

3-(1,1- dimethylethyl)-4- ({5-[(4R)-4-ethyl-4- methyl-2,5-dioxo-1-imidazolidinyl]-2- pyrimidinyl}oxy) benzonitrile 4-[(5-amino-2-pyrimidinyl) oxy]-3-(1,1- dimethylethyl) benzonitrile (Intermediate 163)¹H NMR (400 MHz, DMSO- d₆) δ ppm 8.78 (2H, s), 8.74 (1H, br. s.), 7.86(1H, d), 7.78 (1H, dd), 7.41 (1H, d), 1.74- 1.86 (1H, m), 1.62- 1.74(1H, m), 1.41 (3H, s), 1.33 (9H, s), 0.88 (3H, t) 1.08 min, 394 [M +H]+. 66

3-(1,1- dimethylethyl)-4- ({5-[(4R)-4-ethyl-4- methyl-2,5-dioxo-1-imidazolidinyl]-2- pyridinyl}oxy) benzonitrile 4-[(5-amino-2-pyridinyl)oxy]- 3-(1,1-dimethylethyl) benzonitrile (Intermediate 146) ¹HNMR (400 MHz, DMSO- d₆) δ ppm 8.62 (1H, s), 8.21 (1H, d), 7.96 (1H, dd),7.84 (1H, d), 7.74 (1H, dd), 7.31 (1H, d), 7.21 (1H, d). 1.74- 1.86 (1H,m), 1.62-1.73 (1H, m), 1.41 (3H, s), 1.37 (9H, s), 0.88 (3H, t) 1.13min, 393 [M + H]+.

Example 674-{[4-(4,4-dimethyl-2,5-dioxo-1-imidazolidinyl)phenyl]oxy}-2-(methyloxy)benzonitrile

N¹-(4-{[4-cyano-3-(methyloxy)phenyl]oxy}phenyl)-2-methylalaninamide(Intermediate 191, 77.0 mg) was dissolved in DCM (10 mL). Triethylamine(0.218 mL, 1.57 mmol) was added and the obtained mixture was cooled at0° C. Bis(trichloromethyl)carbonate (68.1 mg, 0.22 mmol) was dissolvedin 5 mL of DCM and the obtained solution was added dropwise to thereaction mixture. The reaction mixture was stirred at 0° C. After 15min, the reaction mixture was evaporated in vacuo to obtain the crudeproduct that was purified by silica gel chromatography (from 100:0 to50:50 Cyclohexane/EtOAc in 10 CV; then 50:50 Cyclohexane/EtOAc for 10CV) to obtain 65.1 mg of the title compound as a white solid

¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.56 (1H, br. s.) 7.72 (1H, d)7.42-7.49 (2H, m) 7.19-7.29 (2H, m) 6.97 (1H, d) 6.57 (1H, dd) 3.89 (3H,s) 1.41 (6H, s); UPLC_ipqc: 0.93 min, 352 [M+H]+.

The following compounds were prepared using the foregoing methodology,replacingN¹-(4-{[4-cyano-3-(methyloxy)phenyl]oxy}phenyl)-2-methylalaninamide(Intermediate 191) with the appropriate amine, as described in theforegoing Reaction Schemes. Final products were purified byflash-chromatography (Silica cartridge; Cyclohexane/EtOAc or otherappropriate solvent system).

UPLC_ipqc characteri- Ex. Structure Name Amine NMR characterizationzation 68

4-{[4-(4,4-dimethyl- 2,5-dioxo-1- imidazolidinyl) phenyl]oxy}-2-(ethyloxy) benzonitrile N¹-(4-{[4-cyano-3- {ethyloxy)phenyl]oxy}phenyl)-2- methylalaninamide (Intermediate 193) ¹H NMR (400 MHz,CDCl₃): δ ppm 7.50 (1H, d) 7.45-7.50 (2H, m) 7.12-7.17 (2H, m) 6.60 (1H,d) 6.55(1H, dd) 5.77 (1H, br. s.) 4.08 (2H, q) 1.58 (6H, s) 1.47 (3H, t)1.00 min, 366 [M + H)+ 69

4-({4-[(4R)-4-ethyl- 2,5-dioxo-1- imidazolidinyl] phenyl}oxy)-2-(ethyloxy) benzonitrile (2R)-2-amino-N- (4-{[4-cyano-3-(ethyloxy)phenyl] oxy}phenyl) butanamide (Intermediate 194) ¹H NMR (400MHz, CDCl₃): δ ppm 7.49 (1H, d), 7.43-7.48 (2H, m), 7.12-7.18 (2H, m),6.61 (1H, d), 6.55 (1H, dd), 5.64 (1H, br. s.), 4.22 (1H, ddd), 4.09(2H, q), 1.98-2.09 (1H, m), 1.88- 1.00 min, 366 [M + H]+. 1.98 (1H, m),1.48 (3H, t), 1.08 (3H, t) 70

3-cyclopropyl-4- ({5-[(4R)-4-ethyl- 2,5-dioxo-1- imidazolidinyl]-2-pyridinyl}oxy) benzonitrile (2R)-2-amino-N- {6-[(4-cyano-2-cyclopropylphenyl) oxy]-3-pyridinyl} butanamide (Intermediate 188) ¹HNMR (400 MHz, DMSO-d₆): δ ppm 8.64 (1H, br. s), 8.14-8.11 (1H, m),7.96-7.84 (1H, m), 7.71-7.66 (1H, m), 7.52 (1H, br. s), 7.29 (2H, d),4.24-4.18 (1H, m), 1.97-1.89 (1H, m), 1.86- 1.78 (1H, m), 1.75-1.67 (1H,m), 0.95 (3H, t), 0.91-0.85 (2H, m), 0.81-0.75 (2H, m) 0.98 min, 363[M + H]⁺. 71

3-(1,1- dimethylethyl)-4- ({5-[(4R)-4-ethyl- 2,5-dioxo-1-imidazolidinyl]-2- pyridinyl}oxy) benzonitrile (2R)-2-amino-N-(6-{[4-cyano-2- (1,1- dimethylethyl) phenyl]oxy}-3- pyridinyl)butanamide (Intermediate 189) ¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.64 (1H,br. s), 8.19 (1H, br. s), 7.96-7.90 (1H, m), 7.82 (1H, br. s), 7.76-7.68 (1H, m). 7.30 (1H, d), 7.19 (1H, d), 4.25- 4.17 (1H, m), 1.86-1.77(1H, m), 1.76-1.66 (1H, m), 1.35 (9H, s), 0.95 (3H, t) 1.10 min, 379[M + H]⁺. 72

4-({5-[(4R)-4-ethyl- 2,5-dioxo-1- imidazolidinyl]-2- pyridinyl}oxy)-2-(methyloxy) benzonitrile (2R)-2-amino-N- (6-{[4-cyano-3-(methyloxy)phenyl] oxy}-3- pyridinyl) butanamide (Intermediate 190) ¹HNMR (400 MHz, CDCl₃): δ ppm 8.30 (1H, dd) 7.85 (1H, dd) 7.58 (1H, dt)7.10 (1H, dd) 6.74-6.84 (2H, m) 6.22 (1H, br. s.) 4.20-4.26 (1H, m) 3.92(3H, s) 1.97-2.08 (1H, m) 1.85-1.97 (1H, m) 1.07 0.87 min, 353 [M + H]+.(3H, t) 73

4-({4-[(4R)-4-ethyl- 2,5-dioxo-1- imidazolidinyl] phenl}oxy)-2-(methyloxy) benzonitrile (2R)-2-amino-N- (4-{[4-cyano-3-(methyloxy)phenyl] oxy}phenyl) butanamide (Intermediate 192) ¹H NMR (400MHz, CDCl₃): δ ppm 7.50 (1H, d) 7.43-7.48 (2H, m) 7.12-7.19 (2H, m) 6.64(1H, d) 6.56 (1H, dd) 6.08 (1H, br. s.) 4.18-4.24 (1H, m) 3.89 (3H, s)1.98-2.08 (1H, m) 1.87-1.98 (1H, m) 1.08 0.94 min, 352 [M + H]+. (3H, t)74

2-[(cyclopropyl- methyl)oxy]-4- ({5-[(4R)- 4-ethyl-2,5-dioxo-1-imidazolidinyl]-2- pyridinyl}oxy) benzonitrile (2R)-2-amino-N-[6-({4-cyano-3- [(cyclopropyl- methyl)oxy] phenyl}oxy)-3- pyridinyl]butanamide (Intermediate 195) ¹H NMR (400 MHz, CDCl₃): δ ppm 8.29 (1H,dd), 7.83 (1H, dd), 7.58 (1H, dd), 7.09 (1H, dd), 6.75-6.79 (2H, m).6.07 (1H, br. s.), 4.23 (1H, ddd), 3.91 (2H, d), 1.97- 2.08 (1H, m),1.86-1.98 (1H, m), 1.25-1.37 (1H, 1.01 min, 393 [M + H]+. m), 1.07 (3H,t), 0.62- 0.72 (2H, m), 0.35-0.46 (2H, m) 75

(5R)-5-ethyl-3-[6- ({4-methyl-3- [(trifluoromethyl) oxy]phenyl}oxy)-3-pyridinyl]-2,4- imidazolidinedione (2R)-2-amino-N- [6-({4-methyl-3-[(trifluoromethyl) oxy]phenyl}oxy)-3- pyridinyl] butanamide(Intermediate 196) ¹H NMR (400 MHz, CDCl₃): δ ppm 8.26 (1H, dd), 7.77(1H, dd), 7.28 (1H, m), 7.05-7.10 (1H, m), 7.00-7.05 (2H, m), 6.36 (1H,br. s.), 4.21 (1H, ddd), 2.32 (3H, s), 1.96-2.05 (1H, m), 1.84-1.96 (1H,m), 1.06 1.14 min, 396 [M + H]+. (3H, t) 76

2-cyclopropyl-4- ({5-[(4R)-4-ethyl- 2,5-dioxo-1- imidazolidinyl]-2-pyridinyl}oxy) benzonitrile (2R)-2-amino-N- {6-[(4-cyano-3-cyclopropylphenyl) oxy]-3-pyridinyl} butanamide (Intermediate 197)¹H-NMR (400 MHz, DMSO-d₆): δ ppm 8.63 (1H, br. s.), 8.19 (1H, d), 7.92(1H, dd), 7.82 (1H, d), 7.25 (1H, d), 7.13 (1H, dd), 6.93 (1H, d), 4.19-4.26 (1H, m), 2.16-2.26 (1H, m), 1.78-1.88 (1H, m), 1.67-1.77 (1H, m),1.09-1.18 (2H, m), 0.97 (3H, t), 0.82-0.90 (2H, 0.99 min, 363 [M + H]+m) 77

4-({5-[(4R)-4-ethyl- 2,5-dioxo-1- imidazolidinyl]-2-pyridinyl}oxy)-2-(1- methylethyl) benzonitrile (2R)-2-amino-N-(6-{[4-cyano-3-(1- methylethyl)phenyl] oxy}-3-pyridinyl) butanamide(Intermediate 198) ¹H-NMR (400 MHz, DMSO-d₆): δ ppm 8.66 (1H, s) 8.19(1H, d) 7.93 (1H, dd) 7.84 (1H, d) 7.36 (1H, d) 7.28 (1H, d) 7.18 (1H,dd) 4.19-4.25 (1H, m) 3.21-3.30 (1H, m) 1.77-1.87 (1H, m) 1.65- 1.76(1H, m) 1.27 (6H, d) 0.96 (3H, t) 1.03 min, 365 [M + H]+ 78

4-({5-[(4R)-4-ethyl- 2,5-dioxo-1- imidazolidinyl]-2-pyridinyl}oxy)-2-(1- methylethyl) benzonitrile (2R)-2-amino-N-{6-[(4-cyano-3- ethylphenyl)oxy]- 3-pyridinyl} butanamide (Intermediate199) ¹H-NMR (400 MHz, DMSO-d₆): δ ppm 8.66 (1H, s) 8.19 (1H, dd) 7.93(1H, dd) 7.85 (1H, d) 7.32 (1H, d) 7.28 (1H, dd) 7.19 (1H, dd) 4.19-4.25(1H, m) 2.82 (2H, q) 1.77- 1.88 (1H, m) 1.65-1.77 (1H, m) 1.23 (3H, t)0.96 (3H, t) 0.98 min, 351 [M + H]+ 79

(5R)-5-ethyl-3-[2- ({4-methyl-3- [(trifluoromethyl) oxy]phenyl}oxy)-5-pyrimidinyl]-2,4- imidazolidinedione (2R)-2-amino-N- [2-({4-methyl-3-[(trifluoromethyl) oxy]phenyl}oxy)-5- pyrimidinyl] butanamide(Intermediate 200) ¹H-NMR (400 MHz, DMSO-d₆): δ ppm 8.76 (1H, br. s.),8.70 (2H, s), 7.48 (1H, d), 7.33-7.37 (1H, m), 7.26 (1H, dd), 4.20-4.27(1H, m), 2.31 (3H, s), 1.77-1.90 (1H, m), 1.65-1.77 (1H, m), 0.98 (3H,t) 1.08 min, 397 [M + H]+ 80

4-({5-[(4R)-4-ethyl- 2,5-dioxo-1- imidazolidinyl]-2- pyridinyl}oxy)-2-[(1-methylethyl) oxy]benzonitrile (2R)-2-amino-N- [6-({4-cyano-3-[(1-methylethyl)oxy] phenyl}oxy)-3- pyridinyl] butanamide (Intermediate 201)¹H-NMR (400 MHz, DMSO-d₆): δ ppm 8.65 (1H, s), 8.20 (1H, d), 7.93 (1H,dd), 7.76 (1H, d), 7.27 (1H, d), 7.16 (1H, d), 6.84 (1H, dd), 4.74-4.85(1H, m), 4.17-4.26 (1H, m), 1.76-1.89 (1H, m), 1.65-1.76 (1H, m), 1.31(6H, d), 0.96 (3H, t) 1.00 min, 381 [M + H]+. 81

4-({5-[(4R)-4-ethyl- 2,5-dioxo-1- imidazolidinyl]-2- pyridinyl}oxy)-3-methylbenzonitrile (2R)-2-amino-N- {6-[(4-cyano-2- methylphenyl)oxy]-3-pyridinyl} butanamide (Intermediate 202) ¹H NMR (400 MHz,DMSO-d₆): δ ppm 8.64 (1H, s), 8.08-8.14 (1H, m), 7.88-7.95 (1H, m), 7.86(1H, s), 7.70-7.77 (1H, m), 7.28 (2H, t), 4.15-4.26 (1H, m), 2.17 (3H,s), 1.62-1.89 (2H, m), 0.95 (3H, t) 0.90 min, 337 [M + H]+ 82

4-({5-[(4R)-4-ethyl- 2,5-dioxo-1- imidazolidinyl]-2- pyridlnyl}oxy)-2-[(trifluoromethyl) oxy]benzonitrile (2R)-2-amino-N- [6-({4-cyano-3-[(trifluoromethyl) oxy]phenyl}oxy)-3- pyridinyl] butanamide(Intermediate 203) ¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.67 (1H, s),8.19-8.26 (1H, m), 8.14 (1H, d), 7.94- 8.02 (1H, m), 7.66 (1H, s),7.43-7.51 (1H, m), 7.36 (1H, d), 4.18-4.27 (1H, m), 1.63-1.91 (2H, m),0.96 (3H, t) 1.02 min, 407 [M + H]+, 405 [M − H]− 83

3-ethyl-4-({5-[(4R)- 4-ethyl-2,5-dioxo- 1-imidazolidinyl]-2-pyrimidinyl}oxy) benzonitrile (2R)-2-amino-N- {2-[(4-cyano-2-ethylphenyl)oxy]-5- pyrimidinyl} butanamide (Intermediate 204) ¹H-NMR(400 MHz, DMSO-d₆): δ ppm 8.78 (1H, s) 8.73 (2H, s) 7.91 (1H, d) 7.79(1H, dd) 7.45 (1H, d) 4.21-4.27 (1H, m) 2.53-2.58 (2H, m) 1.77-1.88 (1H,m) 1.66- 1.76 (1H, m) 1.12 (3H, t) 0.97 (3H, t) 0.93 min, 352 [M + H]+84

4-({5-[(4R)-4-ethyl- 2,5-dioxo-1- imidazolidinyl]-2- pyrimidinyl}oxy)-3-methylbenzonitrile (2R)-2-amino-N- {2-[(4-cyano-2- methylphenyl)oxy]-5-pyrimidinyl} butanamide (Intermediate 205) ¹H-NMR (400 MHz,DMSO-d₆): δ ppm 8.77 (1H, s) 8.73 (2H, s) 7.91 (1H, d) 7.79 (1H, dd)7.45 (1H, d) 4.21-4.27 (1H, m) 2.16 (3H, s) 1.77- 1.88 (1H, m) 1.65-1.77(1H, m) 0.97 (3H, t) 0.85 min, 338 [M + H]+ 85

3-(1,1- dimethylethyl)-4- ({5-[(4R)-4-ethyl- 2,5-dioxo-1-imidazolidinyl]-2- pyrimidinyl}oxy) benzonitrile (2R)-2-amino-N-(2-{[4-cyano-2-(1, 1-dimethylethyl) phenyl]oxy}-5- pyrimidinyl)butanamide (Intermediate 206) ¹H-NMR (400 MHz, DMSO-d₆): δ ppm 8.78 (1H,br. s.), 8.75 (2H, s), 7.86 (1H, d), 7.79 (1H, dd), 7.42 (1H, d), 4.21-4.27 (1H, m), 1.78-1.89 (1H, m), 1.65-1.78 (1H, m), 1.33 (9H, s), 0.98(3H, t) 1.03 min, 380 [M + H]+

Example 864-({5-[(4R)-4-ethyl-4-methyl-2,5-dioxo-1-imidazolidinyl]-2-pyridinyl}oxy)-2-(1-methylethyl)benzonitrile

To a solution of4-({5-[(4R)-4-ethyl-4-methyl-2,5-dioxo-1-imidazolidinyl]-2-pyridinyl}oxy)-2-(1-methylethenyl)benzonitrile(Intermediate 207, 98 mg) in MeOH (10 mL) Pd 10% w/w on activate carbon(10 mg) was added and the reaction mixture was stirred for 1 hour underH₂ atmosphere (P=1 atm). The catalyst was filtered off and the solventremoved under reduced pressure. The residue was purified by flashchromatography on silica gel (SNAP 10 g) eluting from 75:25 to 40:60cyclohexane/ethyl acetate affording the title compound (80 mg) as whitesolid.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.61 (1H, br. s.), 8.21 (1H, d), 7.94(1H, dd), 7.84 (1H, d), 7.35 (1H, d), 7.27 (1H, d), 7.18 (1H, dd),3.21-3.30 (1H, m), 1.73-1.85 (1H, m), 1.60-1.72 (1H, m), 1.40 (3H, s),1.28 (6H, d), 0.88 (3H, t); UPLC_ipqc: 1.08 min, 379 [M+H]+.

Reference Intermediate 208 1,3-bis{[(methyloxy)methyl]oxy}benzene

To a solution of 1,3-benzenediol (1.5 g, 13.62 mmol) in dryN,N-Dimethylformamide (13.62 ml) at 0° C. sodium hydride (0.981 g, 40.9mmol) was added and the reaction mixture was stirred for 15 minutes atthe same temperature. MOM-Cl (3.10 ml, 40.9 mmol) was quickly added andthe reaction mixture was stirred for 1 hour while the temperature wasallowed to reach room temperature. The reaction was quenched with brine(20 ml) and extracted with ethyl acetate (3×50 ml). The organic layerwas washed with brine (2×30 ml), dried over sodium sulphate, filteredand evaporated and the residue was purified by flash chromatography(Biotage system) on silica gel using a 50 g SNAP column and cyclohexaneto cyclohexane/ethyl acetate 8:2 as eluents affording the title compound(1.59 g, 8.02 mmol) as a colourless oil.

¹H NMR (400 MHz, DMSO): δ ppm 7.16-7.23 (1H, d), 6.69-6.64 (3H, m), 5.17(4H, s), 3.38 (6H, s).

Reference Intermediate 209 ethyl(2,6-bis{[(methyloxy)methyl]oxy}phenyl)(oxo)acetate

To a solution of 1,3-bis{[(methyloxy)methyl]oxy}benzene (ReferenceIntermediate 208, 2.19 g) in dry tetrahydrofuran (10 ml) at roomtemperature BuLi 1.6M in hexane (8.29 ml, 13.26 mmol) was added and thereaction mixture was stirred for 30 minutes at the same temperature. Themixture was cooled to −78° C. and it was added (via cannulation) to asolution of ethyl chloro(oxo)acetate (2.263 g, 16.57 mmol) in drytetrahydrofuran (10 ml) at −78° C. The reaction mixture was stirred at−78° C. for 30 minutes. The reaction was quenched with an aqueoussaturated solution of ammonium chloride (10 ml) and extracted with ethylacetate (2×30 ml). Combined organic layers were dried over sodiumsulphate, filtered and evaporated. The residue was purified by flashchromatography (Biotage system) on silica gel using a 100 g SNAP columnand cyclohexane to cyclohexane/ethyl acetate 8:2 as eluent affording thetitle compound as a light yellow oil (1.75 g).

¹H NMR (400 MHz, DMSO): δ ppm 7.46 (1H, t), 6.87 (2H, d), 5.20 (4H, s),4.29 (2H, q), 3.34 (6H, s), 1.27 (3H, t).

Reference Intermediate 210 ethyl2-(2,6-bis{[(methyloxy)methyl]oxy}phenyl)-2-propenoate

To a suspension of methyltriphenylphosphonium bromide (3.13 g, 8.75mmol) in dry tetrahydrofuran (30 ml) at 0° C. KHMDS (1.745 g, 8.75 mmol)was slowly added and the reaction mixture was stirred for 15 minutes at0° C. and for 45 minutes at room temperature. The reaction mixture wascooled to 0° C. and a solution of ethyl(2,6-bis{[(methyloxy)methyl]oxy}phenyl)(oxo)acetate (ReferenceIntermediate 209, 1.74 g) in dry tetrahydrofuran (10 mL) was slowlyadded and the reaction mixture was stirred for 2 hours at 0° C. Thereaction was quenched with an aqueous saturated solution of ammoniumchloride (10 ml), diluted with water (20 ml) and extracted with ethylacetate (2×50 ml). The organic layer was dried over sodium sulphate,filtered and evaporated. The residue was purified by flashchromatography (Biotage system) on silica gel using a 100 g SNAP columnand cyclohexane to cyclohexane/ethyl acetate 8:2 as eluents affordingthe title compound as a colourless oil (1.37 g).

¹H NMR (400 MHz, DMSO-d6): δ ppm 7.21 (1H, t), 6.78 (2H, d), 6.44 (1H,d), 5.74 (1H, d), 5.12 (4H, s), 4.12 (2H, q), 3.32 (6H, s), 1.17 (3H,t).

Reference Intermediate 211 ethyl1-(2,6-bis{[(methyloxy)methyl]oxy}phenyl)cyclopropanecarboxylate

To a solution of trimethylsulfoxonium iodide (1.805 g, 8.20 mmol) in drydimethyl sulfoxide (20 mL) sodium hydride 60% dispersion in mineral oil(0.310 g, 7.75 mmol) was added and the reaction mixture was stirred for1 hour at room temperature. A solution of ethyl2-(2,6-bis{[(methyloxy)methyl]oxy}phenyl)-2-propenoate (ReferenceIntermediate 210, 1.35 g) in dry dimethyl sulfoxide (10 mL) was slowlyadded and the reaction mixture was stirred for 1 hour at roomtemperature. The reaction was quenched with an aqueous saturatedsolution of ammonium chloride (10 ml), diluted with water (20 ml) andextracted with ethyl acetate (2×50 ml). The organic layer was washedwith water (50 ml), dried over sodium sulphate, filtered and evaporated.The residue was purified by flash chromatography (Biotage system) onsilica gel using a 50 g SNAP column and cyclohexane to cyclohexane/ethylacetate 8:2 as eluents affording the title as a colourless oil (1.14 g).

¹H NMR (400 MHz, DMSO): δ ppm 7.15 (1H, t), 6.71 (2H, d), 5.18 (4H, s),3.97 (2H, q), 3.36 (6H, s), 1.53-1.58 (2H, m), 1.09-1.14 (2H, m), 1.04(3H, t).

Reference Intermediate 2122-[1-(hydroxymethyl)cyclopropyl]-3-{[(methyloxy)methyl]oxy}phenol

To a solution of ethyl1-(2,6-bis{[(methyloxy)methyl]oxy}phenyl)cyclopropanecarboxylate(Reference Intermediate 211, 490 mg) in ethanol (10 ml) HCl 2N in water(0.789 mL, 1.579 mmol) was added and the reaction mixture was stirredovernight at 50° C. Toluene (20 mL) was added and the combined solventswere removed under reduced pressure. The residue was re-suspended intoluene (20 ml) and the solvent evaporated. The obtained residue wasdissolved in dry tetrahydrofuran (20 ml), the mixture was cooled to 0°C. and NaH 60% dispersion in mineral oil (126 mg, 3.16 mmol) was addedand the reaction mixture was stirred for 30 minutes at the sametemperature. MOM-Cl (0.120 mL, 1.579 mmol) was then added and thereaction mixture was stirred for 2 hours at 0° C. LiAlH4 (1 M in THF,1.579 ml, 1.579 mmol) was added and the reaction mixture was furtherstirred for 1 hour at the same temperature. The reaction was quenchedwith an aqueous saturated solution of ammonium chloride (10 ml), dilutedwith water (10 ml) and extracted with ethyl acetate (2×50 ml). Combinedorganic layers were dried over sodium sulphate, filtered and evaporatedand the residue was purified by flash chromatography (Biotage system) onsilica gel using a 25 g SNAP column and cyclohexane to cyclohexane/ethylacetate 7:3 as eluents affording the title compound as a colourless oil(191 mg).

¹H NMR (400 MHz, DMSO): δ ppm 8.90 (1H, br.s) 6.96 (1H, t), 6.50 (1H,d), 6.45 (1H, d), 5.16 (2H, s), 4.93 (1H, br.s), 3.45 (2H, s), 3.40 (3H,s), 0.86-0.93 (2H, m), 0.56-0.62 (2H, m); UPLC: 0.59 min, 225 [M+H]+.

Reference Intermediate 2134-{[(methyloxy)methyl]oxy}spiro[1-benzofuran-3,1′-cyclopropane]

To a solution of2-[1-(hydroxymethyl)cyclopropyl]-3-{[(methyloxy)methyl]oxy}phenol(Reference Intermediate 212, 190 mg) in dry tetrahydrofuran (10 ml)triphenylphosphine (333 mg, 1.271 mmol) was added and the reactionmixture was stirred until complete dissolution of PPh3. DIAD (0.198 ml,1.017 mmol) was then added dropwise and the reaction mixture was stirredfor 30 minutes at room temperature The solvent was removed under reducedpressure. The residue was purified by flash chromatography (Biotagesystem) on silica gel using a 25 g SNAP column and cyclohexane tocyclohexane/ethyl acetate 9:1 as eluents affording the title compound asa light yellow oil (120 mg).

¹H NMR (400 MHz, DMSO): δ ppm 6.97 (1H, t), 6.51 (1H, d), 6.43 (1H, d),5.12 (2H, s), 4.40 (2H, s), 3.35 (3H, s), 1.43-1.48 (2H, m), 0.85-0.90(2H, m); UPLC_B: 0.88 min, 207 [M+H]+.

Reference Intermediate 214 spiro[1-benzofuran-3,1′-cyclopropan]-4-ol

To a solution of4-{[(methyloxy)methyl]oxy}spiro[1-benzofuran-3,1′-cyclopropane](Reference Intermediate 213, 118 mg) in methanol (5 ml), HCl 2N in water(0.286 mL, 0.572 mmol) was added and the reaction mixture was stirredovernight at 50° C. Combined solvents were removed under reducedpressure and the residue was re-dissolved in toluen (10 ml) and thesolvent was removed. The residue was purified by flash chromatography(Biotage system) on silica gel using a 10 g SNAP column and cyclohexaneto cyclohexane/ethyl acetate 7:3 as eluents affording the title compoundas a white solid (70 mg).

¹H NMR (400 MHz, DMSO): δ ppm 9.28 (1H, s), 6.81 (1H, t), 6.24 (1H, d),6.22 (1H, d), 4.34 (2H, s), 1.40-1.45 (2H, m), 0.77-0.82 (2H, m).

Reference Intermediate 2155-nitro-2-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)pyridine

To a solution of spiro[1-benzofuran-3,1′-cyclopropan]-4-ol (ReferenceIntermediate 214, 70 mg) in dry N,N-dimethylformamide (2 ml) potassiumcarbonate (89 mg, 0.647 mmol) and then 2-chloro-5-nitropyridine (75 mg,0.475 mmol) were added and the reaction mixture was stirred for 3 hoursat 100° C. The reaction was quenched with brine (1 ml), diluted withwater (2 ml) and extracted with ethyl acetate (3×10 ml). The organiclayer was dried over sodium sulphate, filtered and evaporated. Theresidue was purified by flash chromatography (Biotage system) on silicagel using a 10 g SNAP column and cyclohexane to cyclohexane/ethylacetate 9:1 as eluents affording the title compound as a white solid(100 mg).

¹H NMR (400 MHz, DMSO): δ ppm 9.05 (1H, d), 8.63 (1H, dd), 7.23 (1H, d),7.13 (1H, t), 6.73 (1H, d), 6.60 (1H, d), 4.45 (2H, s), 1.05-1.10 (2H,m), 0.88-0.93 (2H, m);

UPLC: 0.79 min, 285 [M+H]+.

Reference Intermediate 2166-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)-3-pyridinamine

To a solution of5-nitro-2-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)pyridine(Reference Intermediate 215, 99 mg) in tetrahydrofuran (5 ml)/water (2.5ml) iron (97 mg, 1.741 mmol) and then ammonium chloride (93 mg, 1.741mmol) were added and the reaction mixture was stirred for 4 hours atroom temperature. The catalyst was filtered off and the residue wasdiluted with an aqueous saturated solution of NaHCO3 (5 ml) andextracted with ethyl acetate (3×10 ml). The organic layer was dried oversodium sulphate, filtered and evaporated and the residue was purified byflash chromatography (Biotage system) on silica gel using a 10 g SNAPcolumn and cyclohexane/ethyl acetate 8:2 to cyclohexane/ethyl acetate1:1 as eluents affording the title compound as a light yellow solid (85mg).

¹H NMR (400 MHz, DMSO): 5 ppm 7.52 (1H, d), 7.06 (1H, dd), 6.97 (1H, t),6.70 (1H, d), 6.53 (1H, d), 6.23 (1H, d), 5.08 (2H, s), 4.43 (2H, s),1.28-1.33 (2H, m), 0.86-0.91 (2H, m); UPLC: 0.62 min, 255 [M+H]+.

Reference Intermediate 217

1,1-dimethylethyl[(1R)-1-({[6-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)-3-pyridinyl]amino}carbonyl)propyl]carbamate

To a solution of(2R)-2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)butanoic acid (94 mg,0.462 mmol) in dry N,N-Dimethylformamide (2 mL) DIPEA (0.115 mL, 0.661mmol) and then TBTU (159 mg, 0.496 mmol) were added and the reactionmixture was stirred for 15 minutes at room temperature.6-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)-3-pyridinamine(Reference Intermediate 216, 84 mg) was added and the reaction mixturewas stirred for 6 hours at the same temperature. The reaction wasquenched with brine (2 ml), diluted with water (5 ml) and extracted withethyl acetate (2×10 ml). The organic layer was washed with ice coldbrine (2×5 ml), dried over sodium sulphate, filtered and evaporated. Theresidue was purified by flash chromatography (Biotage system) on silicagel using a 10 g SNAP column and cyclohexane to cyclohexane/ethylacetate 7:3 as eluents affording the title compound as a colourless oil(130 mg).

¹H NMR (400 MHz, DMSO): δ ppm 10.14 (1H, br.s), 8.32 (1H, d), 8.08 (1H,dd), 7.02-7.09 (2H, m), 6.96 (1H, d), 6.63 (1H, d), 6.42 (1H, d), 4.44(2H, s), 3.93-4.01 (1H, m), 1.52-1.75 (2H, m), 1.39 (9H, s), 1.15-1.22(2H, m), 0.85-0.95 (5H, m); UPLC: 0.80 min, 440 [M+H]+.

Reference Intermediate 218(2R)-2-amino-N-[6-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)-3-pyridinyl]butanamide

To a solution of 1,1-dimethylethyl[(1R)-1-(({[6-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)-3-pyridinyl]amino}carbonyl)propyl]carbamate(Reference Intermediate 217, 128 mg) in dry dichloromethane (3 ml) at 0°C. TFA (0.9 mL, 11.68 mmol) was slowly added and the reaction mixturewas stirred for 2 hours at the same temperature. The reaction wasdiluted with dichloromethane (10 ml) and an aqueous saturated solutionof NaHCO3 was added while the pH was allowed to reach ˜8. Two phaseswere separated and the aqueous layer was re-extracted withdichloromethane (10 ml). The organic layers were combined, dried oversodium sulphate, filtered and evaporated affording the title compound asa colourless oil (92 mg).

¹H NMR (400 MHz, DMSO): δ ppm 8.37 (1H, d), 8.13 (1H, dd), 7.05 (1H, t),6.95 (1H, d), 6.63 (1H, d), 6.42 (1H, d), 4.44 (2H, s), 3.24 (1H, m),1.61-1.72 (1H, m), 1.44-1.55 (1H, m), 1.16-1.21 (2H, m), 0.91 (3H, t),0.86-0.91 (2H, m); UPLC_B: 0.74 min, 340 [M+H]+.

Reference Intermediate 219 1,1-dimethylethyl(1,1-dimethyl-2-oxo-2-{[6-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)-3-pyridinyl]amino}ethyl)carbamate

To a solution of N-{[(1,1-dimethylethyl)oxy]carbonyl}-2-methylalanine(80 mg, 0.393 mmol) in dry N,N-dimethylformamide (1.5 mL) DIPEA (0.096mL, 0.551 mmol) and then HATU (150 mg, 0.393 mmol) were added and thereaction mixture was stirred for 15 minutes at room temperature Thissolution was added to a solution of6-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)-3-pyridinamine(Reference Intermediate 218, 40 mg) in dry N,N-dimethylformamide (0.5ml) and the reaction mixture was stirred overnight at room temperature.The reaction was quenched with water (2 ml), diluted with brine (10 ml)and extracted with ethyl acetate (2×20 ml). The organic layer was driedover sium sulphqte, filtered and evaporated and the residue was purifiedby flash chromatography (Biotage system) on silica gel using a 10 g SNAPcolumn and cyclohexane/ethyl acetate 8:2 to cyclohexane/ethyl acetate1:1 as eluents affording the title compound as a white solid (52 mg).

¹H NMR (400 MHz, DMSO): δ ppm 9.62 (1H, br.s), 8.24-8.42 (1H, br.m),8.05 (1H, d), 6.98-7.10 (2H, m), 6.92 (1H, d), 6.61 (1H, d), 6.40 (1H,d), 4.44 (2H, s), 1.42 (6H, s), 1.36 (9H, s), 1.15-1.21 (2H, m),0.85-0.91 (2H, m); UPLC: 0.81 min, 440 [M+H]+.

Reference Intermediate 2192-methyl-N¹-[6-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)-3-pyridinyl]alaninamide

To a solution of 1,1-dimethylethyl(1,1-dimethyl-2-oxo-2-({[6-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)-3-pyridinyl]amino}ethyl)carbamate(Reference Intermediate 219, 50 mg) in dry dichloromethane (4 mL) at 0°C. TFA (1 ml, 12.98 mmol) was slowly added and the reaction mixture wasstirred for 2 hours at the same temperature. The reaction was dilutedwith dichloromethane (10 ml) and an aqueous saturated solution of NaHCO3was added while the pH was allowed to reach ˜8. Two phases wereseparated and the organic layer was dried over sodium sulphate, filteredand evaporated affording the title compound (35 mg) as a colourless oil.

¹H NMR (400 MHz, DMSO): δ ppm 8.40 (1H, d), 8.15 (1H, dd), 7.04 (1H, t),6.94 (1H, d), 6.62 (1H, d), 6.41 (1H, d), 4.43 (2H, s), 1.28 (6H, s),1.15-1.20 (2H, m), 0.86-0.91 (2H, m); UPLC: 0.56 min, 340 [M+H]+.

Reference Example 87(5R)-5-ethyl-3-[6-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)-3-pyridinyl]-2,4-imidazolidinedione

To a solution of(2R)-2-amino-N-[6-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)-3-pyridinyl]butanamide(Reference Intermediate 218, 90 mg) in dry dichloromethane (15 ml) TEA(0.185 ml, 1.326 mmol) was added and the reaction mixture was cooled to0° C. A solution of triphosgene (35.4 mg, 0.119 mmol) in drydichloromethane (5 mL) was slowly added and the reaction mixture wasstirred for 30 minutes at the same temperature. The reaction wasquenched with water (10 ml) and two phases were separated. The organiclayer was dried over sodium sulphate, filtered and evaporated. Theresidue was purified by flash chromatography (Biotage system) on silicagel using a 10 g SNAP column and cyclohexane/ethyl acetate 8:2 tocyclohexane/ethyl acetate 1:1 as eluents affording the title compound(65 mg, 0.178 mmol) as a white solid.

¹H NMR (400 MHz, DMSO): 5 ppm 8.63 (1H, s), 8.14 (1H, d), 7.85 (1H, dd),7.11 (1H, s), 7.09 (1H, t), 6.68 (1H, dd), 6.52 (1H, dd), 4.45 (2H, s),4.18-1.24 (1H, m), 1.76-1.88 (1H, m), 1.64-1.76 (1H, m), 1.13-1.18 (2H,m), 0.96 (3H, t), 0.89-0.94 (2H, m); UPLC_B: 0.78 min, 366 [M+H]+.

Reference Example 885,5-dimethyl-3-[6-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)-3-pyridinyl]-2,4-imidazolidinedione

To a solution of2-methyl-N1-[6-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)-3-pyridinyl]alaninamide(Reference Intermediate 219, 34 mg) in dry dichloromethane (6 mL) TEA(0.070 mL, 0.501 mmol) was added and the mixture was cooled to 0° C. Asolution of triphosgene (13.38 mg, 0.045 mmol) in dry dichloromethane (2mL) was slowly added and the reaction mixture was stirred for 1 hour atthe same temperature. The reaction was quenched with water (3 ml) andtwo phases were separated. The organic layer was dried over soiumsulphate, filtered and evaporated. The residue was purified by flashchromatography (Biotage system) on silica gel using a 10 g SNAP columnSNAP and cyclohexane/ethyl acetate 7:3 to cyclohexane/ethyl acetate 3:7as eluents affording the title as a white solid (23 mg).

¹H NMR (400 MHz, DMSO): δ ppm 8.63 (1H, s), 8.17 (1H, d), 7.88 (1H, d),7.06-7.12 (2H, m), 6.67 (1H, d), 6.51 (1H, d), 4.45 (2H, s), 1.41 (6H,s), 1.12-1.17 (2H, m), 0.88-0.93 (2H, m); UPLC: 0.73 min, 366 [M+H]+.

Example 89 Biological Assay

The ability of the compounds of the invention to modulate thevoltage-gated potassium channel subtypes Kv3.2/3.1 may be determinedusing the following assay.

Cell Biology

A stable cell line expressing human Kv3.2 channels (hKv3.2) was createdby transfecting Chinese Hamster Ovary (CHO)-K1 cells with a pClH5-hKv3.2vector. Cells were cultured in DMEM/F12 medium supplemented by 10%Foetal Bovine Serum, 1× non-essential amino acids (Invitrogen) and 500ug/ml of Hygromycin-B (Invitrogen). Cells were grown and maintained at37° C. in a humidified environment containing 5% CO₂ in air.

To assess compound effects on human Kv3.1 channels (hKv3.1),CHO/Gam/E1A-clone22 alias CGE22 cells were transduced using a hKv3.1BacMam reagent. This cell line was designed to be an improvedCHO-K1-based host for enhanced recombinant protein expression ascompared to wild type CHO-K1. The cell line was generated following thetransduction of CHO-K1 cells with a BacMam virus expressing theAdenovirus-Gam1 protein and selection with Geneticin-G418, to generate astable cell line, CHO/Gam-A3. CHO/Gam-A3 cells were transfected withpcDNA3-E1A-Hygro, followed by hygromycin-B selection and FACS sorting toobtain single-cell clones. BacMam-Luciferase and BacMam-GFP viruses werethen used in transient transduction studies to select the clone based onhighest BacMam transduction and recombinant protein expression. CGE22cells were cultured in the same medium used for the hKv3.2 CHO-K1 stablecell line with the addition of 300 ug/ml hygromycin-B and 300 ug/mlG418. All other conditions were identical to those for hKv3.2 CHO-K1cells. The day before an experiment 10 million CGE22 cells were platedin a T175 culture flask and the hKv3.1 BacMam reagent (pFBM/human Kv3.1)was added (MOI of 50). Transduced cells were used 24 hours later.

Cell Preparation for IonWorks Quattro™ Experiments

The day of the experiment, cells were removed from the incubator and theculture medium removed. Cells were washed with 5 ml of Dulbecco's PBS(DPBS) calcium and magnesium free and detached by the addition of 3 mlVersene (Invitrogen, Italy) followed by a brief incubation at 37° C. for5 minutes. The flask was tapped to dislodge cells and 10 ml of DPBScontaining calcium and magnesium was added to prepare a cell suspension.The cell suspension was then placed into a 15 ml centrifuge tube andcentrifuged for 2 min at 1200 rpm. After centrifugation, the supernatantwas removed and the cell pellet re-suspended in 4 ml of DPBS containingcalcium and magnesium using a 5 ml pipette to break up the pellet. Cellsuspension volume was then corrected to give a cell concentration forthe assay of approximately 3 million cells per ml.

All the solutions added to the cells were pre-warmed to 37° C.

Electrophysiology

Experiments were conducted at room temperature using IonWorks Quattro™planar array electrophysiology technology (Molecular Devices Corp.) withPatchPlate™ PPC. Stimulation protocols and data acquisition were carriedout using a microcomputer (Dell Pentium 4). Planar electrode holeresistances (Rp) were determined by applying a 10 mV voltage step acrosseach well. These measurements were performed before cell addition. Aftercell addition and seal formation, a seal test was performed by applyinga voltage step from −80 mV to −70 mV for 160 ms. Following this,amphotericin-B solution was added to the intracellular face of theelectrode to achieve intracellular access. Cells were held at −70 mV.Leak subtraction was conducted in all experiments by applying 50 mshyperpolarizing (10 mV) prepulses to evoke leak currents followed by a20 ms period at the holding potential before test pulses. From theholding potential of −70 mV, a first test pulse to −15 mV was appliedfor 100 ms and following a further 100 ms at −70 mV, a second pulse to40 mV was applied for 50 ms. Cells were then maintained for a further100 ms at −100 mV and then a voltage ramp from −100 mV to 40 mV wasapplied over 200 ms. In all experiments, the test pulses protocol wasperformed in the absence (pre-read) and presence (post-read) of the testcompound. Pre- and post-reads were separated by the compound additionfollowed by a 3 minute incubation.

Solutions and Drugs

The intracellular solution contained the following (in mM): K-gluconate100, KCl 54, MgCl2 3.2, HEPES 5, adjusted to pH 7.3 with KOH.Amphotericin-B solution was prepared as 50 mg/ml stock solution in DMSOand diluted to a final working concentration of 0.1 mg/ml inintracellular solution. The external solution was Dulbecco's PhosphateBuffered Saline (DPBS) and contained the following (in mM): CaCl2 0.90,KCl 2.67, KH2PO4 1.47, MgCl.6H2O 0.493, NaCl 136.9, Na3PO4 8.06, with apH of 7.4.

Compounds of the invention (or reference compounds such asN-cyclohexyl-N-[(7,8-dimethyl-2-oxo-1,2-dihydro-3-quinolinyl)methyl]-N′-phenylureawere dissolved in dimethylsulfoxide (DMSO) at a stock concentration of10 mM. These solutions were further diluted with DMSO using a Biomek FX(Beckman Coulter) in a 384 compound plate. Each dilution (1 μL) wastransferred to another compound plate and external solution containing0.05% pluronic acid (66 μL) was added. 3.5 μL from each plate containinga compound of the invention was added and incubated with the cellsduring the IonWorks Quattro™ experiment. The final assay dilution was200 and the final compound concentrations were in the range 50 μM to 50nM.

Data Analysis

The recordings were analysed and filtered using both seal resistance(>20 MΩ) and peak current amplitude (>500 pA at the voltage step of 40mV) in the absence of compound to eliminate unsuitable cells fromfurther analysis. Paired comparisons between pre- and post-drugadditions measured for the −15 mV voltage step were used to determinethe positive modulation effect of each compound. Data were normalized tothe maximum effect of the reference compound (50 microM ofN-cyclohexyl-N-[(7,8-dimethyl-2-oxo-1,2-dihydro-3-quinolinyl)methyl]-N′-phenylurea)and to the effect of a vehicle control (0.5% DMSO). The normalized datawere analysed using ActivityBase software. The concentration of compoundrequired to increase currents by 50% (pEC50) was determined by fittingof the concentration-response data using a four parameter logisticfunction in ActivityBase.

N-cyclohexyl-N-[(7,8-dimethyl-2-oxo-1,2-dihydro-3-quinolinyl)methyl]-N′-phenylureawas obtained from ASINEX (Registry Number: 552311-06-5).

All the Example compounds were tested in the above assay anddemonstrated potentiation of Kv3.1 or Kv3.2 or Kv3.1 and Kv 3.2 (herein“Kv-3.1 and/or Kv3.2”) whole-cell currents of, on average, at least 20%of that observed with 50 microMN-cyclohexyl-N-[(7,8-dimethyl-2-oxo-1,2-dihydro-3-quinolinyl)methyl]-N′-phenylurea.Thus, in the recombinant cell assays of Example x, all of the Examplesact as positive modulators. As used herein, a Kv3.1 and/or Kv3.2positive modulator is a compound which has been shown to produce atleast 20% potentiation of whole-cell currents mediated by human Kv3.1and/or human Kv3.2 channels recombinantly expressed in mammalian cells,as determined using the assays described in Example 89 (BiologicalAssays).

A secondary analysis of the data from the assays described in Example 89investigates the effect of the compounds on rate of rise of the currentfrom the start of the depolarizing voltage pulses. The magnitude of theeffect of a compound can be determined from the time constant(Tau_(act)) obtained from a non-linear fit, using the equation givenbelow, of the rise in Kv3.1 or Kv3.2 currents following the start of thedepolarizing voltage pulse.

Y═(Y0−Ymax)*exp(−K*X)+Ymax

Where:

-   -   Y0 is the current value at the start of the depolarizing voltage        pulse;    -   Ymax is the plateau current;

K is the rate constant, and Tau_(act) is the activation time constant,which is the reciprocal of K.

Similarly, the effect of the compounds on the time taken for Kv3.1 andKv3.2 currents to decay on closing of the channels at the end of thedepolarizing voltage pulses can also be investigated. In this lattercase, the magnitude of the effect of a compound on channel closing canbe determined from the time constant (Tau_(deact)) of a non-linear fitof the decay of the current (“tail current”) immediately following theend of the depolarizing voltage pulse.

The time constant for activation (Tau_(act)) has been determined forseveral of the compounds of the Examples. FIG. 1 shows the data for twocompounds of the invention. Table 1 provides the Tau_(act) data for allof the Examples analysed in this way.

FIG. 1 a shows hKv3.2 currents recorded using the assay described inExample 89. Data shown are the individual currents over the period ofthe depolarizing voltage step to −15 mV recorded from 4 different cellsat two concentrations of compound (Example 19). The data are fitted by asingle exponential curve (solid lines) using the fitting procedure inPrism version 5 (Graphpad Software Inc).

FIG. 1 b shows hKv3.2 currents recorded using the assay described inExample 89. Data shown are the individual currents over the period ofthe depolarizing voltage step to −15 mV recorded from 2 different cellsat two concentrations of the compound of Example 71. The data are fittedby a single exponential curve (solid lines) using the fitting procedurein Prism version 5 (Graphpad Software Inc).

TABLE 1 Summary hKv3.2 data from the analysis of activation time(Tau_(act)). To allow for comparison between compounds, the compoundconcentration chosen was that which produced a similar current (~0.3 nA)at the end of the voltage pulse, with the exception of the vehicle,where maximum currents were <0.1 nA. Concentration Standard Number ofExample (μM) Tau_(act) mean (ms) Deviation experiments Vehicle — 7.1 1.76 (cells) 19 6.25 9.9 2.2 5 30 12.5 7.3 1.8 4 87 0.2 50.1 7.5 5 88 0.419.3 1.0 4 71 0.2 23.0 6.2 4 77 0.8 9.2 2.3 2 70 3.1 13.0 2.3 2 76 3.17.6 1.9 2 78 3.1 8.2 2.0 2 82 3.1 10.4 2.8 2 80 3.1 9.7 1.0 2

As can be seen from Table 1, Examples 87, 88 and 71 markedly increasethe value of Tau_(act). Whereas the other compounds analysed had nomarked effect on Tau_(act) compared to the vehicle control (DMSO 0.5%).

Kv3.1 and Kv3.2 channels must activate and deactivate very rapidly inorder to allow neurons to fire actions potentials at high frequency(Rudy and McBain, 2001, Trends in Neurosciences 24, 517-526). Slowing ofactivation will delay the onset of action potential repolarization;slowing of deactivation will lead to hyperpolarizing currents thatreduce the excitability of the neuron and delay the time before theneuron can fire a further action potential. Together these two slowingeffects will lead to a reduction rather than a facilitation of theneurons ability to fire at high frequencies. Thus compounds that havethis slowing effect on the Kv3.1 and/or Kv3.2 channels will effectivelybehave as negative modulators of the channels, leading to a slowing ofneuronal firing. This latter effect can be observed from recordings madefrom “fast-firing” interneurons in the cortex of rat brain, usingelectrophysiological techniques, in vitro (FIG. 2).

FIG. 2 shows recordings made from identified “fast-firing” interneuronsin the somatosensory cortex of the mouse. The neurons are induced tofire at high frequencies by trains of high frequency depolarizingcurrent pulses at 100, 200, and 300 Hz. The ability of the neuron tofire an action potential on each pulse is determined. A spikeprobability of 1 on the y-axis of the graph indicates that an actionpotential is generated by the neuron on each of the depolarizing currentpulses. In the absence of drug (closed circles, n=9), the neuronsmaintained a spike probability of 1 up to 300 Hz. However, in thepresence of Example 87 (1 microM; open circles, n=6), the neurons wereunable to follow trains at the highest frequency. * p<0.05, ANOVA forrepeated measures.

Therefore, although all the Examples herein identified act as positivemodulators in the recombinant cell assay of Example 89, those compoundswhich markedly increase the value of Tau_(act) reduce the ability ofneurons in native tissue to fire at high speeds, and as a result act asnegative modulators.

Compounds which act as positive modulators include Examples 19, 30, 77,70, 76, 78, 82 and 80

Compounds which act as negative modulators include Examples 87, 88, and71.

In one aspect of the invention, there is provided a Kv3 potentiatingcompound which is associated with a mean tau value that is not more that2 standard deviations greater than the mean value obtained in thepresence of vehicle (DMSO 0.5%), for use in the treatment of disorderswhere positive modulation of Kv3.1 and/or Kv3.2 channel function isbeneficial, including schizophrenia, bipolar disorder, hearingdisorders, sleep disorders, substance-related disorders, and epilepsy.

In one aspect of the invention, there is provided a Kv3 potentiatingcompound which is associated with a mean tau value that is more that 2standard deviations greater than the mean value obtained in the presenceof vehicle (DMSO 0.5%), for use in the treatment of disorders whereinhibition of Kv3.1 and/or Kv3.2 channel function is beneficial,including hyperacusis, Fragile-X, and autism.

Preclinical Experiments

All in vivo studies were conducted in compliance with Project Licensesobtained according to Italian law (art. 7, Legislative Decree no. 116,27 Jan. 1992), which acknowledged the European Directive 86/609/EEC, andwith the GlaxoSmithKline company policy on the care and use oflaboratory animals and related codes of practice.

In the studies that follow, Compound 19 is the compound of Example 19.

Example 90 Evaluation of Compound Effects on the Firing of Interneuronsin the Somatosensory Cortex of Mice, In Vitro

Animals

Transgenic mice [CB6-Tg (Gad1-EGFP) G42Zjh/J] were purchased from TheJackson Laboratory (Maine, USA). These mice selectively express enhancedgreen fluorescent protein (EGFP) in the calcium-binding proteinparvalbumin (Pv)-expressing subclass of basket interneurons. EGFPexpression is not reported in other interneuron classes positive forsomatostatin (SOM), cholecystokinin (CCK), calretinin (CR), and VIP.These mice are therefore useful for the identification of thePv-expressing subset of GABAergic neurons that express Kv3.1 and Kv3.2channels and are able to fire at high frequency.

Slice Preparation

Experiments were performed on 250-μm-thick brain slices containing thesomatosensory cortex. Briefly, brains were removed from deeplyanaesthetized (isofluorane) 25-35 day-old Gad1-EGFP mice. Slices werecut using a DTK 1000 microslicer (DSK, Japan) in the following solution(in mM): KCl (2.5), CaCl₂ (0.1), NaH₂PO₄ (1.2), MgCl₂ (5), NaHCO₃ (26),sucrose (189) and glucose (10), kept at 2-6° C. and gassed with 95%O₂-5% CO₂. After cutting, the slices were left to equilibrate in arecovery chamber for at least one hour in an artificial cerebrospinalfluid (ACSF) containing (in mM): NaCl (120), KCl (2.5), CaCl₂ (2),NaH₂PO₄ (2.5), MgCl₂ (1.5), NaHCO₃ (26), and glucose (10), at roomtemperature and saturated with 95% O₂-5% CO₂.

Electrophysiological Recordings

For electrophysiological recordings, a slice was transferred to asubmersion chamber mounted on the stage of an upright microscope(Axioskop, Carl Zeiss, Germany) and superfused with oxygenated ACSF.Visualization of neurons in the slices was accomplished with a 40×objective using infrared-differential interference contrast (IR-DIC)video microscopy (Hamamatsu C5985, Hamamatsu City, Japan).Parvalbumin-positive interneurons were identified by illuminating thepreparation with a fluorescence lamp with a GFP-filter and switchingbetween fluorescence and IR-DIC video microscopy. Only GFP-positiveneurons were recorded. Whole-cell recordings were made usingborosilicate-glass patch pipettes pulled using a Sutter P-97 electrodepuller and filled with an internal solution containing (in mM):KGluconate (125), EGTA (10), HEPES (10), MgCl₂ (1), KCl (10) and MgATP(2); pH 7.3 adjusted with KOH. When filled with this internal solution,patch electrodes had a tip resistance of 4-7 MΩ. Recordings were carriedout at room temperature (20-22° C.) using a Multiclamp 700B amplifier(Axon Instruments, Foster City, Calif., USA). Current-command protocols(indicated below) and data acquisition were performed using pClamp 10.0software and a Digidata 1320A interface (Axon Instruments, Foster City,Calif., USA). Capacitive transients were neutralised andseries-resistance was monitored continuously throughout the experiment.If it changed by >20% the cell was discarded. Data were filtered at 3kHz and sampled at 10 kHz.

Drugs

Compounds of the invention were dissolved in DMSO (100%),tetraethylammonium (TEA) and tetrodotoxin (TTX), (both from Sigma,Italy) were dissolved in distilled water and stored at −20° C. untiluse. Drugs were diluted to the final concentration on the day of theexperiment. The highest final concentration of DMSO used was 0.1%.

Experimental Procedure

The firing activity of the recorded interneurons was evaluated byapplying long current steps at different intensities. Thus, after theformation of a giga-seal, the amplifier was switched to current-clampmode, allowing the neuron to reach its resting membrane potential. Anegative current was then injected into the cell in order to obtain aresting potential close to −80 mV. From this condition, step currentinjections (50 pA increments, 600 ms) were applied to elicit actionpotentials. This protocol was repeated at least 2 times for each cell.

Online bridge-balance compensation was carried out and R_(m) value wasmonitored continuously throughout the experiment.

Drug Application

Slices were incubated in the recovery chamber for at least 1 hour in thepresence of either vehicle (0.1% DMSO), TEA (0.5 mM)+0.1% DMSO, or TEA(0.5 mM)+Example 19 (1 or 10 microM). After transfer of a slice to therecording chamber, the same drug condition was maintained by superfusionof the appropriate drugs in the circulating ACSF.

Data Acquisition and Analysis

Raw data were acquired using Clampex 10.0 (Molecular Devices, USA). Datawere analyzed using Clampfit 10.0 software (Molecular Devices, USA). Thefrequency of action potential firing (expressed in Hz) in response tostep current injections was calculated from the number of actionpotentials detected over the 600 ms step current. Values of frequencyobtained for each current step in the same experimental condition and inthe same cell were averaged. Since the threshold to evoke actionpotentials differed from one cell to another, current step intensity wasexpressed as pA from the current threshold for action potentialgeneration, rather than in absolute values.

Action potential half-width was calculated for each action potentialusing Clampfit. The values of the 2^(nd)-5^(th) or the last ten actionpotentials evoked by a non-saturating current step (typically 100-150 pAfrom threshold) were averaged for each experimental condition in eachanalyzed cell.

Statistical Analysis

Statistical differences between the effect of treatments on actionfiring frequency were evaluated using a two-way ANOVA for repetitivemeasurements and, if necessary, post hoc planned comparisons(differences were considered significant where p<0.05). The effect ofdrug treatment on action potential half-width and on the firstderivative amplitude was evaluated using an ANOVA. All statisticalanalyses were conducted using Statistica Software (StatSoft version 8).When appropriate, results were reported as mean±SEM.

Criteria for Data Inclusion/Exclusion

The criteria used to include or exclude a cell from the analysis werebased on accurate current-clamp conditions and the stability of therecording throughout the experiment. Online evaluation allowed theexclusion of a cell when the R_(s) and/or R_(m) values changed by >20%.

Results

Interneurons recorded from slices incubated with 0.5 mM TEA fired at alower maximal frequency in response to step currents compared to neuronsrecorded from control slices (Figure X). This effect was significantlyreversed in slices incubated with TEA (0.5 mM) plus Example 19 at 1 μMor 10 μM (one-way ANOVA for repeated measurements, * p<0.05 with respectto TEA alone).

FIG. 3. The frequency of action potentials recorded fromparvalbumin-positive interneurons in the somatosensory cortex of themouse, evoked by depolarizing current steps (600 ms duration andΔ-increment of 50 pA) after at least 1 hour with either vehicle (0.1%DMSO; filled circles, n=6), TEA (0.5 mM)+0.1% DMSO (open circles, n=7),TEA (0.5 mM)+Example 19 (10; filled triangles, n=9), or TEA (0.5mM)+Example 19 (100; open triangles, n=5). * p<0.05; One-way ANOVA forrepeated measurements.

Furthermore, the action potential half-width and was significantlyincreased in cells recorded from slices incubated with TEA (0.5 mM)compared to control slices (0.1% DMSO) (Figure Y). In slices incubatedwith TEA (0.5 mM) plus Example 19 at 1 μM or 10 μM, the mean actionpotential half-width was significantly decreased by 24% and 36%,respectively, compared to slices incubated with TEA (0.5 mM) only (ANOVAand Dunnett test, * p<0.05, n=9; ** p<0.01, n=5, respectively).

FIG. 4. The half-width of evoked action potentials fromparvalbumin-positive interneurons in the somatosensory cortex of themouse. Prior to recordings, slices were incubated for at least 1 hourwith either vehicle (Control; 0.1% DMSO, n=6), TEA (0.5 mM)+0.1% DMSO(n=7), TEA (0.5 mM)+Example 19 (10; n=9), or TEA (0.5 mM)+EXAMPLE 19(100; n=5). * p<0.05; ** p<0.01, *** p<0.001, ANOVA followed by Dunnetttest.

These results demonstrate the ability of compounds of the invention tomodulate the behaviour of fast-firing interneurons in the mouse brain ina manner consistent with positive modulation of Kv3 channels. Theability to enhance Kv3 function in cortical brain areas is alsoconsistent with the potential of these compounds to treat schizophrenia,bipolar disorder, and epilepsy.

Example 91 Evaluation of Compound Effects on Potassium Currents Recordedfrom Neurons in the Medial Nucleus of the Trapezoid Body in Mice, InVitro

Animals

Male CBA/Ca mice (aged 12-16 days) were used in these experiments (inaccordance with the UK Animals Scientific Procedures Act, 1986). Brainslices containing the medial nucleus of the trapezoid body (MNTB) wereprepared as described previously (Brew and Forsythe, 2005).

Drugs

Chemicals and reagents were purchased from Sigma, (Poole, UK) unlessotherwise noted. EXAMPLE 19 was dissolved in DMSO and diluted in ACSF tothe required concentration.

Electrophysiological Recording

Recordings from identified MNTB neurons were conducted as previouslydescribed (Brew and Forsythe, 2005). Slices was placed in a superfusionchamber on an inverted microscope stage and continuously perfused withgassed (95% O₂-5% CO₂) ACSF at a rate of 1 ml min⁻¹ at room temperature.Whole-cell recordings were made from visually identified MNTB neuronsusing an Axopatch 700B amplifier (Molecular Devices, Union City, Calif.,USA). Patch solution comprised (in mM) potassium gluconate (97.5), KCl(32.5), Hepes (40), EGTA (5), MgCl₂ (1), Na₂phosphocreatin (5), pH 7.2with KOH. Pipettes had resistances of 3-5 MΩ and series resistances were6-10 MΩ (compensated by 70%, 10 μs lag). Access resistance wasfrequently monitored and the recording discarded if increases were morethan 2 MΩ.

Once a whole-cell configuration had been obtained, cells were held at−60 mV prior to application of voltage protocols as follows: cells werestepped from the holding potential to −90 for 700 ms and stepped to −40mV for 25 ms and then a voltage pulse to a range of voltages from −100to +40 mV (10 mV increments) was applied for 220 ms before returning tothe holding potential. Following completion of this protocol, TEA (1 mM)was added to the superfusion medium. After 5 minutes, a second set ofrecordings using the same voltage protocol was carried out. Followingthis, Compound 19 (10 microM) was added to the ACSF, in the continuingpresence of TEA (1 mM), and after a further 5 minutes, a final set ofrecordings with the voltage protocol was made.

Statistical Analysis

Currents evoked by the voltage step to +40 mV were compared across drugtreatments for each cell using an unpaired t-test.

Results

TEA (1 mM) significantly reduced the amplitude of outward, highvoltage-activated potassium currents evoked by voltage steps to +40 mV(FIG. 5). This effect was reversed by the subsequent application ofEXAMPLE 19 (10 microM).

FIG. 5. High-voltage activated potassium currents recorded from visuallyidentified MNTB neurons in the mouse, in vitro. Data shown are the mean(+/−s.d.) of the current amplitude evoked by voltage steps to +40 mVunder different drug conditions. TEA (1 mM), TEA (1 mM)+EXAMPLE 19 (10microM). Statistical analysis was conducted using an unpaired t-test.

These data indicate that compounds of the invention can modulate highvoltage-activated potassium currents (presumed to be mediated by Kv3.1channels; Brew and Forsythe, 2005) in neurons of the MNTB, a region ofthe brainstem that processes auditory information. This result supportsthe utility of compounds of the invention for the treatment of hearingdisorders.

Example 92 Electroshock Seizure Model in Rats Experimental Preparation

Male CD rats (85-130 g) were supplied by Charles River, Italy. Animalswere group housed with free access to food (Standard rodent chow) andwater under a 12 h light/dark cycle (lights on at 0600 h). A period ofat least 5 days between arrival at GSK and the study was allowed in allcases.

Experimental Protocol

Animals were administered a test compound at the appropriate dose, routeand pre-treatment time and returned to their home cage. Testing occurredin a separate room from that used for housing. Testing involveddetermining the threshold for tonic hindlimb extensor seizures using aHugo Sachs Electronik stimulator which delivers a constant current of0.3 second duration, 50 Hz, sinewave form, fully adjustable between 1and 300 mA. Stimuli were delivered via corneal electrodes (Stean T O,Atkins A R, Heidbreder C A, Quinn L P, Trail B K, Upton N. (2005) Br J.Pharmacol. 144(5):628-35). Seizure threshold was determined using the‘up and down’ method of Kimball et al. (1957)(Kimball A W, Burnett W TJr, Doherty D G. (1957) Radiat Res. 7(1):1-12). The first animal testedin each group was stimulated with a current that might be expected to beclose to the threshold for induction of a seizure. If a tonic seizurewas not induced, then the next animal in the group received a stimulus 5mA higher. If a tonic seizure was induced, then the next animal receiveda stimulus 5 mA lower. This is repeated for all animals within thecontrol (vehicle) group. In the case of groups treated with a testcompound steps of 5 to 10 mA were used. At the end of the study, bloodsamples were taken for analysis of the drug concentrations in thiscompartment (n=4/group).

Drugs and Materials

All doses were calculated as base. Sodium valproate was suspended inMethocell 1% (w/v) and dosed via the oral (p.o.) route at 5 mL/kg 1 hourbefore test. Compound 19 was dissolved in DMSO and then suspended inMethocell 1% (w/v) to a final DMSO concentration of 5% (v/v). Compound19 was then dosed p.o. at 5 mL/kg 2 hours before test.

Data Analysis

Induction of seizure is measured as an all-or-nothing effect scored aseither present (+) or absent (0) for each animal. The data for eachtreatment group were recorded as the number of +'s and 0's at eachcurrent level employed and this information was then used to calculatethe CC50 value (current required for 50% of animals to show seizurebehaviour)+standard error of the mean according to the method of Kimballet al. (1957). Drug effects were calculated as the % change in CC50.Significant differences between drug-treated animals and appropriatevehicle treated groups were assessed according to the methods ofLitchfield and Wilcoxon (1949).

Results

Pretreatment with Compound 19 was associated with a significant increasein seizure threshold at both doses tested: At the dose of 30 mg/kg p.o.,Compound 19 produced a 91% increase in seizure threshold, whereas at thedose of 60 mg/kg p.o., the increase in seizure threshold was +218%. Theincrease produced by the higher dose of Compound 19 was similar to theincrease produced by the positive control, sodium valproate at 300 mg/kgp.o. (+258%).

Blood concentrations of Compound 19 measured in satellite animals 2hours after dosing were 5.3 and 9.1 μg/mL following the doses of 30 and60 mg/kg p.o., respectively. These concentrations are equivalent tounbound concentrations in blood of 1.3 and 2.2 μM, respectively, andthus are consistent with concentrations of Compound 19 that produce asignificant increase in Kv3-mediated currents observed in the in vitrorecombinant human Kv3 electrophysiology assay, described above.

Conclusions

These results suggest that Compound 19 has anticonvulsant efficacy, andthat this effect is likely to be mediated by the positive modulation ofKv3 potassium channels.

Example 93 Psychostimulant-Induced Hyperactivity in Mice ExperimentalPreparation

Male CD-1 mice (25-35 g) were supplied by Charles River, Italy. Animalswere group housed with free access to food (Standard rodent chow) andwater under a 12 h light/dark cycle (lights on at 0600 h). A period ofat least 5 days between arrival at GSK and the study was allowed in allcases.

Experimental Protocol

Animals were administered a test compound at the appropriate dose, routeand pre-treatment time, and then returned to their home cage. Testingoccurred in a separate room from that used for housing. Mice weretreated orally (p.o.) with the test compound and placed individuallyinto a Perspex box (length 20.5 cm, width 20.5 cm, height 34 cm) coveredwith a perforated lid. Infrared monitoring sensors were located aroundthe perimeter walls (horizontal sensors). Two additional sensors werelocated 2.5 cm above the floor on opposite sides (vertical sensors).Data were collected and analysed using a VersaMax System (AccuscanInstruments Inc., Columbus, Ohio) which in turn transferred informationto a computer. After 30 minutes of habituation, mice were treated withamphetamine dosed intraperitoneally (i.p.) at 2 mg/kg at 10 mL/kg, andsubsequent locomotor activity in the test arena was assessed over afurther 60 minutes. Locomotor activity was determined as the totaldistance (cm) traveled by each mouse in the test arena over the 60minute test period.

Drugs and Materials

All doses were calculated as base. Clozapine was dissolved in distilledwater and dosed at 3 mg/kg intraperitoneum (i.p.) at 10 mL/kg. Compound19 (10, 30 or 60 mg/kg) or vehicle (HPMC 0.5% w/v, Tween80 0.1% v/v inwater) was administered p.o. at 10 mL/kg. Both clozapine and Compound 19were dosed immediately before placing the animal in the test arena (30minutes before amphetamine administration).

Results

Amphetamine alone produced a large and significant increase in totaldistance traveled. A dose of 30 mg/kg p.o. of Compound 19 significantlyreduced the increase in total distance traveled produced by amphetamine.A higher dose of 60 mg/kg p.o. of Compound 19 further reduced theincrease in locomotor activity induced by amphetamine in a mannersimilar to the positive control, clozapine (3 mg/kg i.p.). Data aresummarized in Table 1.

TABLE 1 Effects of Compound 19 on amphetamine induced hyperlocomotion inthe mouse. Compound 19 was administered p.o. 30 minutes beforeamphetamine (2 mg/kg i.p.). Clozapine was administered i.p. 30 minutesbefore amphetamine (2 mg/kg i.p.). Total distance was assessed over 60minutes starting immediately after amphetamine administration. Data areexpressed as mean ± sem. Data were subjected to one-way analysis ofvariance (ANOVA) followed by Dunnett's test (** = p < 0.01 vsamphetamine treatment alone). Total Distance Travelled Treatment (cm)Vehicle  1049 ± 522** Amphetamine (AMPH) 2.0 mg/kg 16304 ± 3309 AMPH 2mg/kg + Compound 19 10 mg/kg 15267 ± 3166 AMPH 2 mg/kg + Compound 19 30mg/kg   5790 ± 1436** AMPH 2 mg/kg + Compound 19 60 mg/kg  1494 ± 378**AMPH 2 mg/kg + Clozapine 3 mg/kg   932 ± 362**

Conclusions

These results show that Compound 19, at doses similar to those that showanticonvulsant efficacy, is able to prevent hyperactivity induced by thepsychostimulant, amphetamine. Compound 19 and other compounds thatpositively modulate Kv3.1 and/or Kv3.2 channels may thus be useful inthe treatment of disorders associated with hyperactivity, such asbipolar mania, or disruption of the dopamine system, such that may occurin drug dependence, attention deficit hyperactivity disorder (ADHD), orschizophrenia.

Example 94 Behaviour of the Common Marmoset

Central anxiolytic effects of a test compound can be assessed from theability of the compound to reduce the characteristic defensive posturesof marmosets in response to a threatening approach by a human. The testcan also be used to assess the sedative or hypnotic effects of a testcompound from its ability to reduce the number of jumps made by theanimals. The study was based on the method described in Costall, B. etal (1988) Br. J. Pharmac. 95 p475P. Laboratory-bred (GSK SpA, Italy)male and female common marmosets over 2 years of age, weighing 300-500 gwere used in the study. The animals were caged in couples in a housingroom maintained at 25±1° C., 60% humidity, and a 12 hour light/darkcycle (lights on at 0600, with 30 minutes simulated dawn and twilight).Both animals in each pair were involved in the test, which was carriedout with the animals in their home cage.

As there can be variability in the behavioural response betweendifferent marmosets, the “responder” animals were pre-selected to meetthe baseline criteria of at least 10 postures exhibited in the 2 minutestest period following the approach of the human operator.

The postures recorded in the test were those described by Costall et alsupra;

-   -   Genital Presenting (“Tail Posture”):

the animal's back is turned to the observer with elevation of the tailto expose the genital region;

-   -   Scent-Marking:

the animal scent-marks the cage surfaces using circum-anal andcircum-genital scent glands;

-   -   Slit-Stare:

the animal stares at the observer with flattened ear tufts and eyesreduced to “slits”

-   -   Arch-Piloerection:

the animal moves around the cage with arched back and full-bodypiloerection, failing to make eye contact with the observer

The number of jumps from the back of the cage to the cage front providedan index of locomotor activity, which could be used to assess thepotential for a hypnotic effect, sedation, or locomotor stimulationproduced by the test compound.

Drugs and Materials

A single dose of Compound 19 (0.3, 1 or 3 mg/kg) or vehicle (HPMC 0.5%(w/v), Tween80 0.1% (v/v) in water) was administered orally (p.o.) 2hours prior to the test (n=5-6 animals per group).

Results

Compound 19 (1 and 3 mg/kg p.o.) significantly reduced the number ofjumps made by the animals over the 2 minute test period, without anyeffects on postures at any dose, indicative of a sedative or hypnoticeffect. Data are summarized in Table 2.

TABLE 2 Effects of Compound 19 on marmoset behaviour. Compound 19 wasadministered p.o. 2 hours before the test. Data are expressed as mean ±sem. Data were subjected to a one-way analysis of variance (ANOVA)followed by Dunnett's test, comparing each compound dose with therelated vehicle treatment (* = p < 0.05 vs vehicle treated animals; ** =p < 0.01 vs vehicle treated animals). Treatment Number of posturesNumber of Jumps Vehicle 12.2 ± 0.3  22.3 ± 2.5 Compound 19 0.3 mg/kg12.2 ± 1.03 21.3 ± 2.7 Compound 19 1 mg/kg 10.0 ± 1.2   15.3 ± 2.5*Compound 19 3 mg/kg 9.7 ± 1.1  11.5 ± 1.9**

Conclusions

These results suggest that Compound 19 has a hypnotic or sedativeprofile in non-human primates, and thus indicates that Compound 19 andother compounds that positively modulate Kv3.1 and/or Kv3.2 channelscould be useful in the treatment of sleep disorders.

Example 95 Pharmaco-Electroencephalography (phEEG) in the CommonMarmoset

Animals and Surgery

Laboratory bred male (vasectomies) and female common marmosets(Callithrix jacchus) over 2 years of age, weighing 250-500 g were usedin this study. The animals were caged in couples, in a housing roommaintained at 25±1° C., 60% humidity and a 12 hour light/dark cycle(lights on at 0600, with 30 min simulated dawn and twilight). Animalsreceived a standard diet and drinking water ad libitum. Only one animalof each pair was involved in the test, which was carried out with theanimal situated in the home cage.

The effect of compounds of the invention was assessed using telemetricrecording of cortical EEG (ECoG). A multichannel telemetric transmitter(DSI model TL11M2-F40-EET) is implanted intraperitoneally using standardsurgical techniques in anaesthetized marmosets. Recording electrodeswere permanently fixed, with dental cement, to the skull directly incontact with the dura mater through two drilled holes in thefronto-parietal region. Following surgery, animals were housed in pairs(one implanted, one unoperated partner) in their home cage with accessto food and water ad libitum. Animals demonstrated a normal behaviouralrepertoire immediately after recovery from surgery; however, phEEG wasassessed at least 3 weeks later. All in vivo studies were conducted inaccordance with the Italian laws and conformed to GlaxoSmithklineethical standards.

Experimental Procedure

The animals were placed in the nest-boxes in their room cages and EEGtraces were recorded using Dataquest ART software for a 5-min period foreach time-point and analyzed using Spike2 software (CED, UK). Thespectral power in each frequency band was determined for each 2 secepoch during the pre-treatment period and averaged; similarly spectralpower in each band was determined for successive 2 sec epochs of each5-min period of recording following vehicle or drug treatment. Change inthe absolute spectral power, for each of the different bands (delta,theta, alpha and beta) was calculated offline.

Drug treatments were assigned according to a complete crossover design:All treatments were randomly distributed between animals, in separateexperimental sessions, each animal received vehicle and each dose ofdrug, after an appropriate wash-out period.

Six animals were treated orally with EXAMPLE 30 at the doses of 0.3, 1and 3 mg/kg (1 ml/kg) and the EEG traces were recorded at +15, 30, 60,90, 120 and 180 minutes following treatment. EXAMPLE 30 was suspended in12.5% (w/v) aqueous captisol containing 0.1% (w/v) Tween80 and 0.5%(w/v) HPMC.

Data Analysis

Four different frequency bands were considered: delta (1.50-6.00 Hz),theta (6.00-8.00 Hz), alpha (8.00-12.00 Hz) and beta (12.00-30.00 Hz).Values for spectral power in each band at each time point were first logtransformed and then analysed with a mixed effect model with time asfixed effect, the baseline level as covariate, and animal as randomterm. Data are summarized as mean of the percentage changes frombaseline and standard error.

Results

The pharmaco-EEG changes observed in these studies show that, comparedto vehicle, EXAMPLE 30 at the highest dose (3 mg/kg) induced astatistically significant increase of the absolute power in the deltaband between 30 and 120 minutes (p<0.05) and a statistically significantincrease in theta band power at 60 minutes (p<0.05). At the intermediatedose (1 mg/kg) EXAMPLE 30 induced a marginally significant (p<0.10)increase in the absolute power in delta band at 30 minutes and aconcomitant significant reduction in the beta band (p<0.05). Nosignificant effects were observed in the alpha band at any dose ofEXAMPLE 30.

These results suggest that compounds of the invention may have anantipsychotic-like profile, since a similar pattern of EEG changes canbe observed with antipsychotic agents in humans.

1-14. (canceled)
 15. A compound selected from the group consisting of:


16. A compound of formula (I):

wherein, X is CH or N; Y is CH or N; and Z is OCH₃, OCH₂CH₃ or OCF₃;wherein, when Z is OCH₃ then X is CH and Y is CH or N; when Z is OCH₂CH₃then X is N and Y is N; and when Z is OCF₃ then X is N and Y is CH or N.17. The compound according to claim 16, which is:4-{[4-methyl-3-(methyloxy)phenyl]oxy}aniline


18. The compound according to claim 16, which is:6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinamine


19. The compound according to claim 16, which is:2-{[3-(ethyloxy)-4-methylphenyl]oxy}-5-pyrimidinamine


20. The compound according to claim 16, which is:6-({4-methyl-3-[(trifluoromethyl)oxy]phenyl}oxy)-3-pyridinamine


21. The compound according to claim 16, which is:2-({4-methyl-3-[(trifluoromethyl)oxy]phenyl}oxy)-5-pyrimidinamine